2021/12/04 更新

写真a

アベ マナブ
阿部 学
ABE Manabu
所属
脳研究所 生命科学リソース研究センター 准教授
医歯学総合研究科 分子細胞医学専攻 准教授
職名
准教授
外部リンク

学位

  • 医学博士 ( 2001年3月   新潟大学 )

  • 理学修士 ( 1997年3月   新潟大学 )

研究キーワード

  • TARPγi

  • 包括脳ネットワーク

  • C57BL

  • NMDA受容体

  • 6

  • 遺伝子トラップ

  • 神経生物学

  • 遺伝子改変マウス

  • 相同組換え

  • 分子

  • 細胞

  • 神経回路

  • シナプス

  • TARPγ

  • 小脳プルキンエ細胞

  • コンディショナルターゲッティング

  • ナプスターゲティング

  • 接着分子

  • Stargazin

  • 脳高次機能

  • 海馬CA3

  • 記憶・学習

  • 遺伝子組換えマウス

  • コンディショナル組換え系

  • 海馬

  • AMPA受容体

  • 細胞接着分子

  • ES細胞

  • グルタミン酸受容体

  • シナプス可塑性

  • βカテニン

研究分野

  • ライフサイエンス / 神経科学一般

  • ライフサイエンス / 神経科学一般

  • ライフサイエンス / 動物生理化学、生理学、行動学

  • ライフサイエンス / 神経科学一般

  • ライフサイエンス / 実験動物学

  • ライフサイエンス / 分子生物学

▶ 全件表示

経歴(researchmap)

  • 新潟大学   脳研究所   准教授

    2012年1月 - 現在

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  • 新潟大学 脳研究所   Brain Research Institute   助教

    2007年 - 2011年12月

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  • 新潟大学 脳研究所   Brain Research Institute   助手

    2001年 - 2007年

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経歴

  • 新潟大学   脳研究所 生命科学リソース研究センター   准教授

    2012年1月 - 現在

  • 新潟大学   医歯学総合研究科 分子細胞医学専攻   准教授

    2012年1月 - 現在

  • 新潟大学   医歯学総合研究科 医科学専攻   准教授

    2012年1月 - 現在

  • 新潟大学   脳研究所 基礎神経科学部門   助教

    2007年4月 - 2011年12月

  • 新潟大学   脳研究所 基礎神経科学部門   助手

    2001年4月 - 2007年3月

所属学協会

 

論文

  • A discrete neuronal circuit induces a hibernation-like state in rodents 査読 国際誌

    Tohru M. Takahashi, Genshiro A. Sunagawa, Shingo Soya, Manabu Abe, Katsuyasu Sakurai, Kiyomi Ishikawa, Masashi Yanagisawa, Hiroshi Hama, Emi Hasegawa, Atsushi Miyawaki, Kenji Sakimura, Masayo Takahashi, Takeshi Sakurai

    Nature   583 ( 7814 )   109 - 114   2020年6月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:Springer Science and Business Media LLC  

    Hibernating mammals actively lower their body temperature to reduce energy expenditure when facing food scarcity1. This ability to induce a hypometabolic state has evoked great interest owing to its potential medical benefits2,3. Here we show that a hypothalamic neuronal circuit in rodents induces a long-lasting hypothermic and hypometabolic state similar to hibernation. In this state, although body temperature and levels of oxygen consumption are kept very low, the ability to regulate metabolism still remains functional, as in hibernation4. There was no obvious damage to tissues and organs or abnormalities in behaviour after recovery from this state. Our findings could enable the development of a method to induce a hibernation-like state, which would have potential applications in non-hibernating mammalian species including humans.

    DOI: 10.1038/s41586-020-2163-6

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    その他リンク: http://www.nature.com/articles/s41586-020-2163-6

  • GluD1 knockout mice with a pure C57BL/6N background show impaired fear memory, social interaction, and enhanced depressive-like behavior. 査読 国際誌

    Chihiro Nakamoto, Meiko Kawamura, Ena Nakatsukasa, Rie Natsume, Keizo Takao, Masahiko Watanabe, Manabu Abe, Tomonori Takeuchi, Kenji Sakimura

    PloS one   15 ( 2 )   e0229288   2020年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    The GluD1 gene is associated with susceptibility for schizophrenia, autism, depression, and bipolar disorder. However, the function of GluD1 and how it is involved in these conditions remain elusive. In this study, we generated a Grid1 gene-knockout (GluD1-KO) mouse line with a pure C57BL/6N genetic background and performed several behavioral analyses. Compared to a control group, GluD1-KO mice showed no significant anxiety-related behavioral differences, evaluated using behavior in an open field, elevated plus maze, a light-dark transition test, the resident-intruder test of aggression and sensorimotor gating evaluated by the prepulse inhibition test. However, GluD1-KO mice showed (1) higher locomotor activity in the open field, (2) decreased sociability and social novelty preference in the three-chambered social interaction test, (3) impaired memory in contextual, but not cued fear conditioning tests, and (4) enhanced depressive-like behavior in a forced swim test. Pharmacological studies revealed that enhanced depressive-like behavior in GluD1-KO mice was restored by the serotonin reuptake inhibitors imipramine and fluoxetine, but not the norepinephrine transporter inhibitor desipramine. In addition, biochemical analysis revealed no significant difference in protein expression levels, such as other glutamate receptors in the synaptosome and postsynaptic densities prepared from the frontal cortex and the hippocampus. These results suggest that GluD1 plays critical roles in fear memory, sociability, and depressive-like behavior.

    DOI: 10.1371/journal.pone.0229288

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  • Non-coding cis-element of Period2 is essential for maintaining organismal circadian behaviour and body temperature rhythmicity. 査読 国際誌

    Masao Doi, Hiroyuki Shimatani, Yuta Atobe, Iori Murai, Hida Hayashi, Yukari Takahashi, Jean-Michel Fustin, Yoshiaki Yamaguchi, Hiroshi Kiyonari, Nobuya Koike, Kazuhiro Yagita, Choogon Lee, Manabu Abe, Kenji Sakimura, Hitoshi Okamura

    Nature communications   10 ( 1 )   2563 - 2563   2019年6月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Non-coding cis-regulatory elements are essential determinants of development, but their exact impacts on behavior and physiology in adults remain elusive. Cis-element-based transcriptional regulation is believed to be crucial for generating circadian rhythms in behavior and physiology. However, genetic evidence supporting this model is based on mutations in the protein-coding sequences of clock genes. Here, we report generation of mutant mice carrying a mutation only at the E'-box cis-element in the promoter region of the core clock gene Per2. The Per2 E'-box mutation abolishes sustainable molecular clock oscillations and renders circadian locomotor activity and body temperature rhythms unstable. Without the E'-box, Per2 messenger RNA and protein expression remain at mid-to-high levels. Our work delineates the Per2 E'-box as a critical nodal element for keeping sustainable cell-autonomous circadian oscillation and reveals the extent of the impact of the non-coding cis-element in daily maintenance of animal locomotor activity and body temperature rhythmicity.

    DOI: 10.1038/s41467-019-10532-2

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  • Rational Engineering of XCaMPs, a Multicolor GECI Suite for In Vivo Imaging of Complex Brain Circuit Dynamics. 査読 国際誌

    Inoue M, Takeuchi A, Manita S, Horigane SI, Sakamoto M, Kawakami R, Yamaguchi K, Otomo K, Yokoyama H, Kim R, Yokoyama T, Takemoto-Kimura S, Abe M, Okamura M, Kondo Y, Quirin S, Ramakrishnan C, Imamura T, Sakimura K, Nemoto T, Kano M, Fujii H, Deisseroth K, Kitamura K, Bito H

    Cell   177 ( 5 )   1346 - 1360   2019年5月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    DOI: 10.1016/j.cell.2019.04.007

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  • Microglia permit climbing fiber elimination by promoting GABAergic inhibition in the developing cerebellum. 査読 国際誌

    Hisako Nakayama, Manabu Abe, Chie Morimoto, Tadatsune Iida, Shigeo Okabe, Kenji Sakimura, Kouichi Hashimoto

    Nature communications   9 ( 1 )   2830 - 2830   2018年7月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Circuit refinement during postnatal development is finely regulated by neuron-neuron interactions. Recent studies suggest participation of microglia in this process but it is unclear how microglia cooperatively act with neuronal mechanisms. To examine roles of microglia, we ablate microglia by microglia-selective deletion of colony-stimulating factor 1 receptor (Csf1r) by crossing floxed-Csf1r and Iba1-iCre mice (Csf1r-cKO). In Csf1r-cKO mice, refinement of climbing fiber (CF) to Purkinje cell (PC) innervation after postnatal day 10 (P10)-P12 is severely impaired. However, there is no clear morphological evidence suggesting massive engulfment of CFs by microglia. In Csf1r-cKO mice, inhibitory synaptic transmission is impaired and CF elimination is restored by diazepam, which suggests that impairment of CF elimination is caused by a defect of GABAergic inhibition on PCs, a prerequisite for CF elimination. These results indicate that microglia primarily promote GABAergic inhibition and secondarily facilitate the mechanism for CF elimination inherent in PCs.

    DOI: 10.1038/s41467-018-05100-z

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  • Fis1 ablation in the male germline disrupts mitochondrial morphology and mitophagy, and arrests spermatid maturation. 国際誌

    Grigor Varuzhanyan, Mark S Ladinsky, Shun-Ichi Yamashita, Manabu Abe, Kenji Sakimura, Tomotake Kanki, David C Chan

    Development (Cambridge, England)   148 ( 16 )   2021年8月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Male germline development involves choreographed changes to mitochondrial number, morphology and organization. Mitochondrial reorganization during spermatogenesis was recently shown to require mitochondrial fusion and fission. Mitophagy, the autophagic degradation of mitochondria, is another mechanism for controlling mitochondrial number and physiology, but its role during spermatogenesis is largely unknown. During post-meiotic spermatid development, restructuring of the mitochondrial network results in packing of mitochondria into a tight array in the sperm midpiece to fuel motility. Here, we show that disruption of mouse Fis1 in the male germline results in early spermatid arrest that is associated with increased mitochondrial content. Mutant spermatids coalesce into multinucleated giant cells that accumulate mitochondria of aberrant ultrastructure and numerous mitophagic and autophagic intermediates, suggesting a defect in mitophagy. We conclude that Fis1 regulates mitochondrial morphology and turnover to promote spermatid maturation.

    DOI: 10.1242/dev.199686

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  • Ddx20, DEAD box helicase 20, is essential for the differentiation of oligodendrocyte and maintenance of myelin gene expression

    Anna Simankova, Norihisa Bizen, Sei Saitoh, Shinsuke Shibata, Nobuhiko Ohno, Manabu Abe, Kenji Sakimura, Hirohide Takebayashi

    Glia   2021年7月

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    掲載種別:研究論文(学術雑誌)   出版者・発行元:Wiley  

    DOI: 10.1002/glia.24058

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    その他リンク: https://onlinelibrary.wiley.com/doi/full-xml/10.1002/glia.24058

  • A comparative analysis of kainate receptor GluK2 and GluK5 knockout mice in a pure genetic background. 国際誌

    Izumi Iida, Kohtarou Konno, Rie Natsume, Manabu Abe, Masahiko Watanabe, Kenji Sakimura, Miho Terunuma

    Behavioural brain research   405   113194 - 113194   2021年5月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Kainate receptors (KARs) are members of the glutamate receptor family that regulate synaptic function in the brain. Although they are known to be associated with psychiatric disorders, how they are involved in these disorders remains unclear. KARs are tetrameric channels assembled from a combination of GluK1-5 subunits. Among these, GluK2 and GluK5 subunits are the major heteromeric subunits in the brain. To determine the functional similarities and differences between GluK2 and GluK5 subunits, we generated GluK2 KO and GluK5 KO mice on a C57BL/6N background, a well-characterized inbred strain, and compared their behavioral phenotypes. We found that GluK2 KO and GluK5 KO mice exhibited the same phenotypes in many tests, such as reduced locomotor activity, impaired motor function, and enhanced depressive-like behavior. No change was observed in motor learning, anxiety-like behavior, or sociability. Additionally, we identified subunit-specific phenotypes, such as reduced motivation toward their environment in GluK2 KO mice and an enhancement in the contextual memory in GluK5 KO mice. These results revealed that GluK2 and GluK5 subunits not only function in a coordinated manner but also have a subunit-specific role in regulating behavior. To summarize, we demonstrated subunit-specific and common behavioral effects of GluK2 and GluK5 subunits for the first time. Moreover, to the best of our knowledge, this is the first evidence of the involvement of the GluK5 subunit in the expression of depressive-like behavior and contextual memory, which strongly indicates its role in psychiatric disorders.

    DOI: 10.1016/j.bbr.2021.113194

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  • CAPS2 deficiency impairs the release of the social peptide, oxytocin, as well as oxytocin-associated social behavior. 国際誌

    Shuhei Fujima, Ryosuke Yamaga, Haruka Minami, Shota Mizuno, Yo Shinoda, Tetsushi Sadakata, Manabu Abe, Kenji Sakimura, Yoshitake Sano, Teiichi Furuichi

    The Journal of neuroscience : the official journal of the Society for Neuroscience   41 ( 20 )   4524 - 4535   2021年4月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Ca2+-dependent activator protein for secretion 2 (CAPS2) regulates dense-core vesicle (DCV) exocytosis to facilitate peptidergic and catecholaminergic transmitter release. CAPS2 deficiency in mice has mild neuronal effects but markedly impairs social behavior. Rare de novo Caps2 alterations also occur in autism spectrum disorder, although whether CAPS2-mediated release influences social behavior remains unclear. Here, we demonstrate that CAPS2 is associated with DCV exocytosis-mediated release of the social interaction modulatory peptide oxytocin (OXT). CAPS2 is expressed in hypothalamic OXT neurons and localizes to OXT nerve projection and OXT release sites, such as the pituitary. Caps2 KO mice exhibited reduced plasma albeit increased hypothalamic and pituitary OXT levels, indicating insufficient release. OXT neuron-specific Caps2 conditional KO supported CAPS2 function in pituitary OXT release, also affording impaired social interaction and recognition behavior that could be ameliorated by exogenous OXT administered intranasally. Thus, CAPS2 appears critical for OXT release, thereby being associated with social behavior.Significance StatementThe role of the neuropeptide, oxytocin, in enhancing social interaction and social bonding behavior has attracted considerable public and neuroscientific attention. A central issue in oxytocin biology concerns how oxytocin release is regulated. Our study provides an important insight into the understanding of oxytocin-dependent social behavior from the perspective of the CAPS2-regulated release mechanism.

    DOI: 10.1523/JNEUROSCI.3240-20.2021

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  • Activity-induced secretion of semaphorin 3A mediates learning. 国際誌

    Aoi Jitsuki-Takahashi, Susumu Jitsuki, Naoya Yamashita, Meiko Kawamura, Manabu Abe, Kenji Sakimura, Akane Sano, Fumio Nakamura, Yoshio Goshima, Takuya Takahashi

    The European journal of neuroscience   2021年3月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    The semaphorin family is a well-characterized family of secreted or membrane-bound proteins that are involved in activity-independent neurodevelopmental processes, such as axon guidance, cell migration, and immune functions. Although semaphorins have recently been demonstrated to regulate activity-dependent synaptic scaling, their roles in Hebbian synaptic plasticity as well as learning and memory remain poorly understood. Here, using a rodent model, we found that an inhibitory avoidance task, a hippocampus-dependent contextual learning paradigm, increased secretion of semaphorin 3A in the hippocampus. Furthermore, the secreted semaphorin 3A in the hippocampus mediated contextual memory formation likely by driving AMPA receptors into hippocampal synapses via the neuropilin1-plexin A4-semaphorin receptor complex. This signaling process involves alteration of the phosphorylation status of collapsin response mediator protein 2, which has been characterized as a downstream molecule in semaphorin signaling. These findings implicate semaphorin family as a regulator of Hebbian synaptic plasticity and learning.

    DOI: 10.1111/ejn.15210

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  • Induction of Mutant Sik3Sleepy Allele in Neurons in Late Infancy Increases Sleep Need. 国際誌

    Kanako Iwasaki, Tomoyuki Fujiyama, Shinya Nakata, Minjeong Park, Chika Miyoshi, Noriko Hotta-Hirashima, Aya Ikkyu, Miyo Kakizaki, Fumihiro Sugiyama, Seiya Mizuno, Manabu Abe, Kenji Sakimura, Satoru Takahashi, Hiromasa Funato, Masashi Yanagisawa

    The Journal of neuroscience : the official journal of the Society for Neuroscience   41 ( 12 )   2733 - 2746   2021年3月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Sleep is regulated in a homeostatic manner. Sleep deprivation increases sleep need, which is compensated mainly by increased EEG δ power during non-rapid eye movement sleep (NREMS) and, to a lesser extent, by increased sleep amount. Although genetic factors determine the constitutive level of sleep need and sleep amount in mice and humans, the molecular entity behind sleep need remains unknown. Recently, we found that a gain-of-function Sleepy (Slp) mutation in the salt-inducible kinase 3 (Sik3) gene, which produces the mutant SIK3(SLP) protein, leads to an increase in NREMS EEG δ power and sleep amount. Since Sik3Slp mice express SIK3(SLP) in various types of cells in the brain as well as multiple peripheral tissues from the embryonic stage, the cell type and developmental stage responsible for the sleep phenotype in Sik3Slp mice remain to be elucidated. Here, we generated two mouse lines, synapsin1CreERT2 and Sik3ex13flox mice, which enable inducible Cre-mediated, conditional expression of SIK3(SLP) in neurons on tamoxifen administration. Administration of tamoxifen to synapsin1CreERT2 mice during late infancy resulted in higher recombination efficiency than administration during adolescence. SIK3(SLP) expression after late infancy increased NREMS and NREMS δ power in male synapsin1CreERT2; Sik3ex13flox/+ mice. The expression of SIK3(SLP) after adolescence led to a higher NREMS δ power without a significant change in NREMS amounts. Thus, neuron-specific expression of SIK3(SLP) after late infancy is sufficient to increase sleep.SIGNIFICANCE STATEMENT The propensity to accumulate sleep need during wakefulness and to dissipate it during sleep underlies the homeostatic regulation of sleep. However, little is known about the developmental stage and cell types involved in determining the homeostatic regulation of sleep. Here, we show that Sik3Slp allele induction in mature neurons in late infancy is sufficient to increase non-rapid eye movement sleep amount and non-rapid eye movement sleep δ power. SIK3 signaling in neurons constitutes an intracellular mechanism to increase sleep.

    DOI: 10.1523/JNEUROSCI.1004-20.2020

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  • Ezh1 regulates expression of Cpg15/Neuritin in mouse cortical neurons.

    Shun Utsunomiya, Yusuke Kishi, Masafumi Tsuboi, Daichi Kawaguchi, Yukiko Gotoh, Manabu Abe, Kenji Sakimura, Kazuma Maeda, Hiroshi Takemoto

    Drug discoveries & therapeutics   15 ( 2 )   55 - 65   2021年3月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Immature neurons undergo morphological and physiological maturation in order to establish neuronal networks. During neuronal maturation, a large number of genes change their transcriptional levels, and these changes may be mediated by chromatin modifiers. In this study, we found that the level of Ezh1, a component of Polycomb repressive complex 2 (PRC2), increases during neuronal maturation in mouse neocortical culture. In addition, conditional knockout of Ezh1 in post-mitotic excitatory neurons leads to downregulation of a set of genes related to neuronal maturation. Moreover, the locus encoding Cpg15/Neuritin (Nrn1), which is regulated by neuronal activity and implicated in stabilization and maturation of excitatory synapses, is a direct target of Ezh1 in cortical neurons. Together, these results suggest that elevated expression of Ezh1 contributes to maturation of cortical neurons.

    DOI: 10.5582/ddt.2021.01017

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  • A Discrete Glycinergic Neuronal Population in the Ventromedial Medulla That Induces Muscle Atonia during REM Sleep and Cataplexy in Mice. 国際誌

    Shuntaro Uchida, Shingo Soya, Yuki C Saito, Arisa Hirano, Keisuke Koga, Makoto Tsuda, Manabu Abe, Kenji Sakimura, Takeshi Sakurai

    The Journal of neuroscience : the official journal of the Society for Neuroscience   41 ( 7 )   1582 - 1596   2021年2月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    During rapid eye movement (REM) sleep, anti-gravity muscle tone and bodily movements are mostly absent, because somatic motoneurons are inhibited by descending inhibitory pathways. Recent studies showed that glycine/GABA neurons in the ventromedial medulla (VMM; GlyVMM neurons) play an important role in generating muscle atonia during REM sleep (REM-atonia). However, how these REM-atonia-inducing neurons interconnect with other neuronal populations has been unknown. In the present study, we first identified a specific subpopulation of GlyVMM neurons that play an important role in induction of REM-atonia by virus vector-mediated tracing in male mice in which glycinergic neurons expressed Cre recombinase. We found these neurons receive direct synaptic input from neurons in several brain stem regions, including glutamatergic neurons in the sublaterodorsal tegmental nucleus (SLD; GluSLD neurons). Silencing this circuit by specifically expressing tetanus toxin light chain (TeTNLC) resulted in REM sleep without atonia. This manipulation also caused a marked decrease in time spent in cataplexy-like episodes (CLEs) when applied to narcoleptic orexin-ataxin-3 mice. We also showed that GlyVMM neurons play an important role in maintenance of sleep. This present study identified a population of glycinergic neurons in the VMM that are commonly involved in REM-atonia and cataplexy.SIGNIFICANCE STATEMENT We identified a population of glycinergic neurons in the ventral medulla that plays an important role in inducing muscle atonia during rapid eye movement (REM) sleep. It sends axonal projections almost exclusively to motoneurons in the spinal cord and brain stem except to those that innervate extraocular muscles, while other glycinergic neurons in the same region also send projections to other regions including monoaminergic nuclei. Furthermore, these neurons receive direct inputs from several brainstem regions including glutamatergic neurons in the sublaterodorsal tegmental nucleus (SLD). Genetic silencing of this pathway resulted in REM sleep without atonia and a decrease of cataplexy when applied to narcoleptic mice. This work identified a neural population involved in generating muscle atonia during REM sleep and cataplexy.

    DOI: 10.1523/JNEUROSCI.0688-20.2020

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  • Dysregulation of sphingolipid metabolic enzymes leads to high levels of sphingosine-1-phosphate and ceramide in human hepatocellular carcinoma. 国際誌

    Kohei Miura, Masayuki Nagahashi, Pankaj Prasoon, Yuki Hirose, Takashi Kobayashi, Jun Sakata, Manabu Abe, Kenji Sakimura, Yasunobu Matsuda, Ali L Butash, Eriko Katsuta, Kazuaki Takabe, Toshifumi Wakai

    Hepatology research : the official journal of the Japan Society of Hepatology   51 ( 5 )   614 - 626   2021年2月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    AIM: Sphingosine-1-phosphate (S1P) and ceramide are bioactive sphingolipids known to be important in regulating numerous processes involved in cancer progression. The aim of this study was to determine the absolute levels of sphingolipids in hepatocellular carcinoma (HCC) utilizing data obtained from surgical specimens. In addition, we explored the clinical significance of S1P in patients with HCC and the biological role of S1P in HCC cells. METHODS: Tumors and normal liver tissues were collected from 20 patients with HCC, and sphingolipids were measured by mass spectrometry. The Cancer Genome Atlas (TCGA) cohort was utilized to evaluate gene expression of enzymes related to sphingolipid metabolism. Immunohistochemistry of phospho-sphingosine kinase 1 (SphK1), an S1P-producing enzyme, was performed for 61 surgical specimens. CRISPR/Cas9-mediated SphK1 knockout cells were used to examine HCC cell biology. RESULTS: S1P levels were substantially higher in HCC tissue compared with normal liver tissue. Levels of other sphingolipids upstream of S1P in the metabolic cascade, such as sphingomyelin, monohexosylceramide and ceramide, were also considerably higher in HCC tissue. Enzymes involved in generating S1P and its precursor, ceramide, were found in higher levels in HCC compared with normal liver tissue. Immunohistochemical analysis found that phospho-SphK1 expression was associated with tumor size. Finally, in vitro assays indicated that S1P is involved in the aggressiveness of HCC cells. CONCLUSIONS: Sphingolipid levels, including S1P and ceramide, were elevated in HCC compared with surrounding normal liver tissue. Our findings suggest S1P plays an important role in HCC tumor progression, and further examination is warranted.

    DOI: 10.1111/hepr.13625

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  • Activin a Receptor Type 2A Mutation Affects the Tumor Biology of Microsatellite Instability-High Gastric Cancer. 国際誌

    Kizuki Yuza, Masayuki Nagahashi, Hiroshi Ichikawa, Takaaki Hanyu, Masato Nakajima, Yoshifumi Shimada, Takashi Ishikawa, Jun Sakata, Shiho Takeuchi, Shujiro Okuda, Yasunobu Matsuda, Manabu Abe, Kenji Sakimura, Kazuaki Takabe, Toshifumi Wakai

    Journal of gastrointestinal surgery : official journal of the Society for Surgery of the Alimentary Tract   2021年1月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    BACKGROUND: Activin A receptor type 2A (ACVR2A) is one of the most frequently mutated genes in microsatellite instability-high (MSI-H) gastric cancer. However, the clinical relevance of the ACVR2A mutation in MSI-H gastric cancer patients remains unclear. The aims of this study were to explore the effect of ACVR2A mutation on the tumor behavior and to identify the clinicopathological characteristics of gastric cancer patients with ACVR2A mutations. METHODS: An in vitro study was performed to investigate the biological role of ACVR2A via CRISPR/Cas9-mediated ACVR2A knockout MKN74 human gastric cancer cells. One hundred twenty-four patients with gastric cancer were retrospectively analyzed, and relations between MSI status, ACVR2A mutations, and clinicopathological factors were evaluated. RESULTS: ACVR2A knockout cells showed less aggressive tumor biology than mock-transfected cells, displaying reduced proliferation, migration, and invasion (P < 0.05). MSI mutations were found in 10% (13/124) of gastric cancer patients, and ACVR2A mutations were found in 8.1% (10/124) of patients. All ACVR2A mutations were accompanied by MSI. The 5-year overall survival rates of ACVR2A wild-type patients and ACVR2A-mutated patients were 57% and 90%, respectively (P = 0.048). Multivariate analysis revealed that older age (P = 0.015), distant metastasis (P < 0.001), and ACVR2A wild-type status (P = 0.040) were independent prognostic factors for overall survival. CONCLUSIONS: Our study demonstrated that gastric cancer patients with ACVR2A mutation have a significantly better prognosis than those without. Dysfunction of ACVR2A in MKN74 human gastric cancer cells caused less aggressive tumor biology, indicating the importance of ACVR2A in the progression of MSI-H tumors.

    DOI: 10.1007/s11605-020-04889-9

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  • Global knockdown of glutamate decarboxylase 67 elicits emotional abnormality in mice. 国際誌

    Shigeo Miyata, Toshikazu Kakizaki, Kazuyuki Fujihara, Hideru Obinata, Touko Hirano, Junichi Nakai, Mika Tanaka, Shigeyoshi Itohara, Masahiko Watanabe, Kenji F Tanaka, Manabu Abe, Kenji Sakimura, Yuchio Yanagawa

    Molecular brain   14 ( 1 )   5 - 5   2021年1月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Reduced expression of glutamate decarboxylase 67 (GAD67), encoded by the Gad1 gene, is a consistent finding in postmortem brains of patients with several psychiatric disorders, including schizophrenia, bipolar disorder and major depressive disorder. The dysfunction of GAD67 in the brain is implicated in the pathophysiology of these psychiatric disorders; however, the neurobiological consequences of GAD67 dysfunction in mature brains are not fully understood because the homozygous Gad1 knockout is lethal in newborn mice. We hypothesized that the tetracycline-controlled gene expression/suppression system could be applied to develop global GAD67 knockdown mice that would survive into adulthood. In addition, GAD67 knockdown mice would provide new insights into the neurobiological impact of GAD67 dysfunction. Here, we developed Gad1tTA/STOP-tetO biallelic knock-in mice using Gad1STOP-tetO and Gad1tTA knock-in mice, and compared them with Gad1+/+ mice. The expression level of GAD67 protein in brains of Gad1tTA/STOP-tetO mice treated with doxycycline (Dox) was decreased by approximately 90%. The GABA content was also decreased in the brains of Dox-treated Gad1tTA/STOP-tetO mice. In the open-field test, Dox-treated Gad1tTA/STOP-tetO mice exhibited hyper-locomotor activity and decreased duration spent in the center region. In addition, acoustic startle responses were impaired in Dox-treated Gad1tTA/STOP-tetO mice. These results suggest that global reduction in GAD67 elicits emotional abnormalities in mice. These GAD67 knockdown mice will be useful for elucidating the neurobiological mechanisms of emotional abnormalities, such as anxiety symptoms associated with psychiatric disorders.

    DOI: 10.1186/s13041-020-00713-2

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  • Heterogeneity of microglial proton channel in different brain regions and its relationship with aging. 国際誌

    Takafumi Kawai, Keizo Takao, Sharmin Akter, Manabu Abe, Kenji Sakimura, Tsuyoshi Miyakawa, Yasushi Okamura

    Journal of neurochemistry   2021年1月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    The properties of microglia largely differ depending on aging as well as on brain regions. However, there are few studies that investigated the functional importance of such heterogeneous properties of microglia at the molecular level. Voltage-gated proton channel, Hv1/VSOP, could be one of the candidates which confers functional heterogeneity among microglia since it regulates brain oxidative stress in age-dependent manner. In this study, we found that Hv1/VSOP shows brain region-dependent heterogeneity of gene expression with the highest level in the striatum. We studied the importance of Hv1/VSOP in two different brain regions, the cerebral cortex and striatum, and examined their relationship with aging (using mice of different ages). In the cortex, we observed the age-dependent impact of Hv1/VSOP on oxidative stress, microglial morphology, and gene expression profile. On the other hand, we found that the age-dependent significance of Hv1/VSOP was less obvious in the striatum than the cortex. Finally, we performed a battery of behavioral experiments on Hv1/VSOP-deficient mice both at young and aged stages to examine the effect of aging on Hv1/VSOP function. Hv1/VSOP-deficient mice specifically showed a marked difference in behavior in light/dark transition test only at aged stages, indicating that anxiety state is altered in aged Hv1/VSOP mice. This study suggests that a combination of brain region heterogeneity and animal aging underscores the functional importance of Hv1/VSOP in microglia.

    DOI: 10.1111/jnc.15292

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  • p62/SQSTM1-droplet serves as a platform for autophagosome formation and anti-oxidative stress response. 国際誌

    Shun Kageyama, Sigurdur Runar Gudmundsson, Yu-Shin Sou, Yoshinobu Ichimura, Naoki Tamura, Saiko Kazuno, Takashi Ueno, Yoshiki Miura, Daisuke Noshiro, Manabu Abe, Tsunehiro Mizushima, Nobuaki Miura, Shujiro Okuda, Hozumi Motohashi, Jin-A Lee, Kenji Sakimura, Tomoyuki Ohe, Nobuo N Noda, Satoshi Waguri, Eeva-Liisa Eskelinen, Masaaki Komatsu

    Nature communications   12 ( 1 )   16 - 16   2021年1月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Autophagy contributes to the selective degradation of liquid droplets, including the P-Granule, Ape1-complex and p62/SQSTM1-body, although the molecular mechanisms and physiological relevance of selective degradation remain unclear. In this report, we describe the properties of endogenous p62-bodies, the effect of autophagosome biogenesis on these bodies, and the in vivo significance of their turnover. p62-bodies are low-liquidity gels containing ubiquitin and core autophagy-related proteins. Multiple autophagosomes form on the p62-gels, and the interaction of autophagosome-localizing Atg8-proteins with p62 directs autophagosome formation toward the p62-gel. Keap1 also reversibly translocates to the p62-gels in a p62-binding dependent fashion to activate the transcription factor Nrf2. Mice deficient for Atg8-interaction-dependent selective autophagy show that impaired turnover of p62-gels leads to Nrf2 hyperactivation in vivo. These results indicate that p62-gels are not simple substrates for autophagy but serve as platforms for both autophagosome formation and anti-oxidative stress.

    DOI: 10.1038/s41467-020-20185-1

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  • Kv11 (ether-à-go-go-related gene) voltage-dependent K+ channels promote resonance and oscillation of subthreshold membrane potentials. 国際誌

    Toshinori Matsuoka, Miwako Yamasaki, Manabu Abe, Yukiko Matsuda, Hiroyuki Morino, Hideshi Kawakami, Kenji Sakimura, Masahiko Watanabe, Kouichi Hashimoto

    The Journal of physiology   599 ( 2 )   547 - 569   2021年1月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    KEY POINTS: Some ion channels are known to behave as inductors and make up the parallel resonant circuit in the plasma membrane of neurons, which enables neurons to respond to current inputs with a specific frequency (so-called 'resonant properties'). Here, we report that heterologous expression of mouse Kv11 voltage-dependent K+ channels generate resonance and oscillation at depolarized membrane potentials in HEK293 cells; expressions of individual Kv11 subtypes generate resonance and oscillation with different frequency properties. Kv11.3-expressing HEK293 cells exhibited transient conductance changes that opposed the current changes induced by voltage steps; this probably enables Kv11 channels to behave like an inductor. The resonance and oscillation of inferior olivary neurons were impaired at the resting membrane potential in Kv11.3 knockout mice. This study helps to elucidate basic ion channel properties that are crucial for the frequency responses of neurons. ABSTRACT: The plasma membranes of some neurons preferentially respond to current inputs with a specific frequency, and output as large voltage changes. This property is called resonance, and is thought to be mediated by ion channels that show inductor-like behaviour. However, details of the candidate ion channels remain unclear. In this study, we mainly focused on the functional roles of Kv11 potassium (K+ ) channels, encoded by ether-á-go-go-related genes, in resonance in mouse inferior olivary (IO) neurons. We transfected HEK293 cells with long or short splice variants of Kv11.1 (Merg1a and Merg1b) or Kv11.3, and examined membrane properties using whole-cell recording. Transfection with Kv11 channels reproduced resonance at membrane potentials depolarized from the resting state. Frequency ranges of Kv11.3-, Kv11.1(Merg1b)- and Kv11.1(Merg1a)-expressing cells were 2-6 Hz, 2-4 Hz, and 0.6-0.8 Hz, respectively. Responses of Kv11.3 currents to step voltage changes were essentially similar to those of inductor currents in the resistor-inductor-capacitor circuit. Furthermore, Kv11 transfections generated membrane potential oscillations. We also confirmed the contribution of HCN1 channels as a major mediator of resonance at more hyperpolarized potentials by transfection into HEK293 cells. The Kv11 current kinetics and properties of Kv11-dependent resonance suggested that Kv11.3 mediated resonance in IO neurons. This finding was confirmed by the impairment of resonance and oscillation at -30 to -60 mV in Kcnh7 (Kv11.3) knockout mice. These results suggest that Kv11 channels have important roles in inducing frequency-dependent responses in a subtype-dependent manner from resting to depolarized membrane potentials.

    DOI: 10.1113/JP280342

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  • Activation of proprotein convertase in the mouse habenula causes depressive-like behaviors through remodeling of extracellular matrix. 国際誌

    Hikaru Ito, Kanako Nozaki, Kenji Sakimura, Manabu Abe, Shigeto Yamawaki, Hidenori Aizawa

    Neuropsychopharmacology : official publication of the American College of Neuropsychopharmacology   46 ( 2 )   442 - 454   2021年1月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    The lateral habenula (LHb) attracts a growing interest as a regulator of monoaminergic activity which were frequently reported to be defective in depression. Here we found that chronic social defeat stress (CSDS) increased production of pro-inflammatory cytokines in LHb associated with mobilization of monocytes and remodeling of extracellular matrix by increased matrix metalloproteinase (MMP) activity. RNA-seq analysis identified proprotein convertase Pcsk5 as an upstream regulator of MMP activation, with upregulation in LHb neurons of mice with susceptibility to CSDS. PCSK5 facilitated motility of microglia in vitro by converting inactive pro-MMP14 and pro-MMP2 to their active forms, highlighting its role in mobilization of microglia and monocytes in neuroinflammation. Suppression of Pcsk5 expression via small interfering RNA (siRNA) ameliorated depressive-like behaviors and pathological mobilization of monocytes in mice with susceptibility to CSDS. PCSK5-MMPs signaling pathway could be a target for development of the antidepressants targeting the inflammatory response in specific brain regions implicated in depression.

    DOI: 10.1038/s41386-020-00843-0

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  • Specific Neuroligin3-αNeurexin1 signaling regulates GABAergic synaptic function in mouse hippocampus. 国際誌

    Motokazu Uchigashima, Kohtarou Konno, Emily Demchak, Amy Cheung, Takuya Watanabe, David G Keener, Manabu Abe, Timmy Le, Kenji Sakimura, Toshikuni Sasaoka, Takeshi Uemura, Yuka Imamura Kawasawa, Masahiko Watanabe, Kensuke Futai

    eLife   9   2020年12月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Synapse formation and regulation require signaling interactions between pre- and postsynaptic proteins, notably cell adhesion molecules (CAMs). It has been proposed that the functions of neuroligins (Nlgns), postsynaptic CAMs, rely on the formation of trans-synaptic complexes with neurexins (Nrxns), presynaptic CAMs. Nlgn3 is a unique Nlgn isoform that localizes at both excitatory and inhibitory synapses. However, Nlgn3 function mediated via Nrxn interactions is unknown. Here we demonstrate that Nlgn3 localizes at postsynaptic sites apposing vesicular glutamate transporter 3-expressing (VGT3+) inhibitory terminals and regulates VGT3+ inhibitory interneuron-mediated synaptic transmission in mouse organotypic slice cultures. Gene expression analysis of interneurons revealed that the αNrxn1+AS4 splice isoform is highly expressed in VGT3+ interneurons as compared with other interneurons. Most importantly, postsynaptic Nlgn3 requires presynaptic αNrxn1+AS4 expressed in VGT3+ interneurons to regulate inhibitory synaptic transmission. Our results indicate that specific Nlgn-Nrxn signaling generates distinct functional properties at synapses.

    DOI: 10.7554/eLife.59545

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  • Central dopamine D2 receptors regulate plasma glucose levels in mice through autonomic nerves. 国際誌

    Hiroko Ikeda, Naomi Yonemochi, Risa Mikami, Manabu Abe, Meiko Kawamura, Rie Natsume, Kenji Sakimura, John L Waddington, Junzo Kamei

    Scientific reports   10 ( 1 )   22347 - 22347   2020年12月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Recent evidence suggests that the central nervous system (CNS) regulates plasma glucose levels, but the underlying mechanism is unclear. The present study investigated the role of dopaminergic function in the CNS in regulation of plasma glucose levels in mice. I.c.v. injection of neither the dopamine D1 receptor agonist SKF 38393 nor the antagonist SCH 23390 influenced plasma glucose levels. In contrast, i.c.v. injection of both the dopamine D2 receptor agonist quinpirole and the antagonist l-sulpiride increased plasma glucose levels. Hyperglycemia induced by quinpirole and l-sulpiride was absent in dopamine D2 receptor knockout mice. I.c.v. injection of quinpirole and l-sulpiride each increased mRNA levels of hepatic glucose-6-phosphatase and phosphoenolpyruvate carboxykinase, which are the key enzymes for hepatic gluconeogenesis. Systemic injection of the β2 adrenoceptor antagonist ICI 118,551 inhibited hyperglycemia induced by l-sulpiride, but not by quinpirole. In contrast, hyperglycemia induced by quinpirole, but not by l-sulpiride, was inhibited by hepatic vagotomy. These results suggest that stimulation of central dopamine D2 receptors increases plasma glucose level by increasing hepatic glucose production through parasympathetic nerves, whereas inhibition of central dopamine D2 receptors increases plasma glucose level by increasing hepatic glucose production through sympathetic nerves.

    DOI: 10.1038/s41598-020-79292-0

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  • AUTS2 Governs Cerebellar Development, Purkinje Cell Maturation, Motor Function and Social Communication. 国際誌

    Kunihiko Yamashiro, Kei Hori, Esther S K Lai, Ryo Aoki, Kazumi Shimaoka, Nariko Arimura, Saki F Egusa, Asami Sakamoto, Manabu Abe, Kenji Sakimura, Takaki Watanabe, Naofumi Uesaka, Masanobu Kano, Mikio Hoshino

    iScience   23 ( 12 )   101820 - 101820   2020年12月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Autism susceptibility candidate 2 (AUTS2), a risk gene for autism spectrum disorders (ASDs), is implicated in telencephalon development. Because AUTS2 is also expressed in the cerebellum where defects have been linked to ASDs, we investigated AUTS2 functions in the cerebellum. AUTS2 is specifically localized in Purkinje cells (PCs) and Golgi cells during postnatal development. Auts2 conditional knockout (cKO) mice exhibited smaller and deformed cerebella containing immature-shaped PCs with reduced expression of Cacna1a. Auts2 cKO and knock-down experiments implicated AUTS2 participation in elimination and translocation of climbing fiber synapses and restriction of parallel fiber synapse numbers. Auts2 cKO mice exhibited behavioral impairments in motor learning and vocal communications. Because Cacna1a is known to regulate synapse development in PCs, it suggests that AUTS2 is required for PC maturation to elicit normal development of PC synapses and thus the impairment of AUTS2 may cause cerebellar dysfunction related to psychiatric illnesses such as ASDs.

    DOI: 10.1016/j.isci.2020.101820

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  • Regulation of hepatic oxidative stress by voltage-gated proton channels (Hv1/VSOP) in Kupffer cells and its potential relationship with glucose metabolism. 国際誌

    Takafumi Kawai, Kento Kayama, Shoki Tatsumi, Sharmin Akter, Nana Miyawaki, Yoshifumi Okochi, Manabu Abe, Kenji Sakimura, Hiroyasu Yamamoto, Shinji Kihara, Yasushi Okamura

    FASEB journal : official publication of the Federation of American Societies for Experimental Biology   34 ( 12 )   15805 - 15821   2020年12月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Voltage-gated proton channels (Hv1/VSOP), encoded by Hvcn1, are important regulator of reactive oxygen species (ROS) production in many types of immune cells. While in vitro studies indicate that Hv1/VSOP regulates ROS production by maintaining pH homeostasis, there are few studies investigating the functional importance of Hv1/VSOP in vivo. In the present study, we first show that Hv1/VSOP is functionally expressed in liver resident macrophage, Kupffer cells, regulating the hepatic oxidative stress in vivo. Our immunocytochemistry and electrophysiology data showed that Hvcn1 is specifically expressed in Kupffer cells, but not in hepatocytes. Furthermore, Hvcn1-deficiency drastically altered the hepatic oxidative stress. The Hvcn1-deficient mice showed high blood glucose and serum insulin but normal insulin sensitivity, indicating that these phenotypes were not linked to insulin resistance. Transcriptome analysis indicated that the gene expression of glycogen phosphorylase (Pygl) and Glucose-6-phosphatase, catalytic subunit (G6pc) were upregulated in Hvcn1-deficient liver tissues, and quantitative PCR confirmed the result for Pygl. Furthermore, we observed higher amount of glucose-6-phosphate, a key sugar intermediate for glucose in Hvcn1-deficient liver than WT, suggesting that glucose production in liver is accelerated in Hvcn1-deficient mice. The present study sheds light on the functional importance of Kupffer cells in hepatic oxidative stress and its potential relationship with glucose metabolism.

    DOI: 10.1096/fj.202001056RRR

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  • Disruption of dystonin in Schwann cells results in late-onset neuropathy and sensory ataxia. 査読 国際誌

    Masao Horie, Nozomu Yoshioka, Satoshi Kusumi, Hiromi Sano, Masayuki Kurose, Izumi Watanabe-Iida, Ibrahim Hossain, Satomi Chiken, Manabu Abe, Kensuke Yamamura, Kenji Sakimura, Atsushi Nambu, Masahiro Shibata, Hirohide Takebayashi

    Glia   68 ( 11 )   2330 - 2344   2020年11月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Dystonin (Dst) is a causative gene for Dystonia musculorum (dt) mice, which is an inherited disorder exhibiting dystonia-like movement and ataxia with sensory degeneration. Dst is expressed in a variety of tissues, including the central nervous system and the peripheral nervous system (PNS), muscles, and skin. However, the Dst-expressing cell type(s) for dt phenotypes have not been well characterized. To address the questions whether the disruption of Dst in Schwann cells induces movement disorders and how much impact does it have on dt phenotypes, we generated Dst conditional knockout (cKO) mice using P0-Cre transgenic mice and Dst gene trap mice. First, we assessed the P0-Cre transgene-dependent Cre recombination using tdTomato reporter mice and then confirmed the preferential tdTomato expression in Schwann cells. In the Dst cKO mice, Dst mRNA expression was significantly decreased in Schwann cells, but it was intact in most of the sensory neurons in the dorsal root ganglion. Next, we analyzed the phenotype of Dst cKO mice. They exhibited a normal motor phenotype during juvenile periods, and thereafter, started exhibiting an ataxia. Behavioral tests and electrophysiological analyses demonstrated impaired motor abilities and slowed motor nerve conduction velocity in Dst cKO mice, but these mice did not manifest dystonic movements. Electron microscopic observation of the PNS of Dst cKO mice revealed significant numbers of hypomyelinated axons and numerous infiltrating macrophages engulfing myelin debris. These results indicate that Dst is important for normal PNS myelin organization and Dst disruption in Schwann cells induces late-onset neuropathy and sensory ataxia. MAIN POINTS: Dystonin (Dst) disruption in Schwann cells results in late-onset neuropathy and sensory ataxia. Dst in Schwann cells is important for normal myelin organization in the peripheral nervous system.

    DOI: 10.1002/glia.23843

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  • Hypnotic effect of thalidomide is independent of teratogenic ubiquitin/proteasome pathway. 国際誌

    Yuki Hirose, Tomohiro Kitazono, Maiko Sezaki, Manabu Abe, Kenji Sakimura, Hiromasa Funato, Hiroshi Handa, Kaspar E Vogt, Masashi Yanagisawa

    Proceedings of the National Academy of Sciences of the United States of America   117 ( 37 )   23106 - 23112   2020年9月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Thalidomide exerts its teratogenic and immunomodulatory effects by binding to cereblon (CRBN) and thereby inhibiting/modifying the CRBN-mediated ubiquitination pathway consisting of the Cullin4-DDB1-ROC1 E3 ligase complex. The mechanism of thalidomide's classical hypnotic effect remains largely unexplored, however. Here we examined whether CRBN is involved in the hypnotic effect of thalidomide by generating mice harboring a thalidomide-resistant mutant allele of Crbn (Crbn YW/AA knock-in mice). Thalidomide increased non-REM sleep time in Crbn YW/AA knock-in homozygotes and heterozygotes to a similar degree as seen in wild-type littermates. Thalidomide similarly depressed excitatory synaptic transmission in the cortical slices obtained from wild-type and Crbn YW/AA homozygous knock-in mice without affecting GABAergic inhibition. Thalidomide induced Fos expression in vasopressin-containing neurons of the supraoptic nucleus and reduced Fos expression in the tuberomammillary nuclei. Thus, thalidomide's hypnotic effect seems to share some downstream mechanisms with general anesthetics and GABAA-activating sedatives but does not involve the teratogenic CRBN-mediated ubiquitin/proteasome pathway.

    DOI: 10.1073/pnas.1917701117

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  • AUTS2 Regulation of Synapses for Proper Synaptic Inputs and Social Communication. 査読 国際誌

    Kei Hori, Kunihiko Yamashiro, Taku Nagai, Wei Shan, Saki F Egusa, Kazumi Shimaoka, Hiroshi Kuniishi, Masayuki Sekiguchi, Yasuhiro Go, Shoji Tatsumoto, Mitsuyo Yamada, Reika Shiraishi, Kouta Kanno, Satoshi Miyashita, Asami Sakamoto, Manabu Abe, Kenji Sakimura, Masaki Sone, Kazuhiro Sohya, Hiroshi Kunugi, Keiji Wada, Mitsuhiko Yamada, Kiyofumi Yamada, Mikio Hoshino

    iScience   23 ( 6 )   101183 - 101183   2020年6月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:Elsevier BV  

    Impairments in synapse development are thought to cause numerous psychiatric disorders. Autism susceptibility candidate 2 (AUTS2) gene has been associated with various psychiatric disorders, such as autism and intellectual disabilities. Although roles for AUTS2 in neuronal migration and neuritogenesis have been reported, its involvement in synapse regulation remains unclear. In this study, we found that excitatory synapses were specifically increased in the Auts2-deficient primary cultured neurons as well as Auts2 mutant forebrains. Electrophysiological recordings and immunostaining showed increases in excitatory synaptic inputs as well as c-fos expression in Auts2 mutant brains, suggesting that an altered balance of excitatory and inhibitory inputs enhances brain excitability. Auts2 mutant mice exhibited autistic-like behaviors including impairments in social interaction and altered vocal communication. Together, these findings suggest that AUTS2 regulates excitatory synapse number to coordinate E/I balance in the brain, whose impairment may underlie the pathology of psychiatric disorders in individuals with AUTS2 mutations.

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  • GRP receptor and AMPA receptor cooperatively regulate itch-responsive neurons in the spinal dorsal horn. 査読 国際誌

    Norikazu Kiguchi, Daisuke Uta, Huiping Ding, Hitoshi Uchida, Fumihiro Saika, Shinsuke Matsuzaki, Yohji Fukazawa, Manabu Abe, Kenji Sakimura, Mei-Chuan Ko, Shiroh Kishioka

    Neuropharmacology   170   108025 - 108025   2020年6月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Gastrin-releasing peptide (GRP) receptor-expressing (GRPR)+ neurons have a central role in the spinal transmission of itch. Because their fundamental regulatory mechanisms are not yet understood, it is important to determine how such neurons are excited and integrate itch sensation. In this study, we investigated the mechanisms for the activation of itch-responsive GRPR+ neurons in the spinal dorsal horn (SDH). GRPR+ neurons expressed the α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptor (AMPAR) containing the GluR2 subunit. In mice, peripherally elicited histaminergic and non-histaminergic itch was prevented by intrathecal (i.t.) administration of the AMPAR antagonist NBQX, which was consistent with the fact that firing of GRPR+ neurons in SDH under histaminergic and non-histaminergic itch was completely blocked by NBQX, but not by the GRPR antagonist RC-3095. Because GRP+ neurons in SDH contain glutamate, we investigated the role of GRP+ (GRP+/Glu+) neurons in regulating itch. Chemogenetic inhibition of GRP+ neurons suppressed both histaminergic and non-histaminergic itch without affecting the mechanical pain threshold. In nonhuman primates, i.t. administration of NBQX also attenuated peripherally elicited itch without affecting the thermal pain threshold. In a mouse model of diphenylcyclopropenone (DCP)-induced contact dermatitis, GRP, GRPR, and AMPAR subunits were upregulated in SDH. DCP-induced itch was prevented by either silencing GRP+ neurons or ablation of GRPR+ neurons. Altogether, these findings demonstrate that GRP and glutamate cooperatively regulate GRPR+ AMPAR+ neurons in SDH, mediating itch sensation. GRP-GRPR and the glutamate-AMPAR system may play pivotal roles in the spinal transmission of itch in rodents and nonhuman primates.

    DOI: 10.1016/j.neuropharm.2020.108025

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  • Generation of Lungs by Blastocyst Complementation in Apneumic Fgf10-Deficient Mice. 査読 国際誌

    Akihiko Kitahara, Qingsong Ran, Kanako Oda, Akihiro Yasue, Manabu Abe, Xulu Ye, Toshikuni Sasaoka, Masanori Tsuchida, Kenji Sakimura, Yoichi Ajioka, Yasuo Saijo, Qiliang Zhou

    Cell reports   31 ( 6 )   107626 - 107626   2020年5月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    The shortage of donor lungs hinders lung transplantation, the only definitive option for patients with end-stage lung disease. Blastocyst complementation enables the generation of transplantable organs from pluripotent stem cells (PSCs) in animal models. Pancreases and kidneys have been generated from PSCs by blastocyst complementation in rodent models. Here, we report the generation of lungs using mouse embryonic stem cells (ESCs) in apneumic Fgf10 Ex1mut/Ex3mutmice by blastocyst complementation. Complementation with ESCs enables Fgf10-deficient mice to survive to adulthood without abnormalities. Both the generated lung alveolar parenchyma and the interstitial portions, including vascular endothelial cells, vascular and parabronchial smooth muscle cells, and connective tissue, largely originate from the injected ESCs. These data suggest that Fgf10 Ex1mut/Ex3mutblastocysts provide an organ niche for lung generation and that blastocyst complementation could be a viable approach for generating whole lungs.

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  • Expression mapping, quantification, and complex formation of GluD1 and GluD2 glutamate receptors in adult mouse brain. 査読 国際誌

    Chihiro Nakamoto, Kohtarou Konno, Taisuke Miyazaki, Ena Nakatsukasa, Rie Natsume, Manabu Abe, Meiko Kawamura, Yugo Fukazawa, Ryuichi Shigemoto, Miwako Yamasaki, Kenji Sakimura, Masahiko Watanabe

    The Journal of comparative neurology   528 ( 6 )   1003 - 1027   2020年4月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    In the cerebellum, GluD2 is exclusively expressed in Purkinje cells, where it regulates synapse formation and regeneration, synaptic plasticity, and motor learning. Delayed cognitive development in humans with GluD2 gene mutations suggests extracerebellar functions of GluD2. However, extracerebellar expression of GluD2 and its relationship with that of GluD1 are poorly understood. GluD2 mRNA and protein were widely detected, with relatively high levels observed in the olfactory glomerular layer, medial prefrontal cortex, cingulate cortex, retrosplenial granular cortex, olfactory tubercle, subiculum, striatum, lateral septum, anterodorsal thalamic nucleus, and arcuate hypothalamic nucleus. These regions were also enriched for GluD1, and many individual neurons coexpressed the two GluDs. In the retrosplenial granular cortex, GluD1 and GluD2 were selectively expressed at PSD-95-expressing glutamatergic synapses, and their coexpression on the same synapses was shown by SDS-digested freeze-fracture replica labeling. Biochemically, GluD1 and GluD2 formed coimmunoprecipitable complex formation in HEK293T cells and in the cerebral cortex and hippocampus. We further estimated the relative protein amount by quantitative immunoblotting using GluA2/GluD2 and GluA2/GluD1 chimeric proteins as standards for titration of GluD1 and GluD2 antibodies. Intriguingly, the relative amount of GluD2 was almost comparable to that of GluD1 in the postsynaptic density fraction prepared from the cerebral cortex and hippocampus. In contrast, GluD2 was overwhelmingly predominant in the cerebellum. Thus, we have determined the relative extracerebellar expression of GluD1 and GluD2 at regional, neuronal, and synaptic levels. These data provide a molecular-anatomical basis for possible competitive and cooperative interactions of GluD family members at synapses in various brain regions.

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  • Decreased Proliferation in the Neurogenic Niche, Disorganized Neuroblast Migration, and Increased Oligodendrogenesis in Adult Netrin-5-Deficient Mice. 国際誌

    Shunsuke Ikegaya, Yurika Iga, Sumiko Mikawa, Li Zhou, Manabu Abe, Kenji Sakimura, Kohji Sato, Satoru Yamagishi

    Frontiers in neuroscience   14   570974 - 570974   2020年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    In the adult mouse brain, neurogenesis occurs mainly in the ventricular-subventricular zone (V-SVZ) and the subgranular zone of the hippocampal dentate gyrus. Neuroblasts generated in the V-SVZ migrate to the olfactory bulb via the rostral migratory stream (RMS) in response to guidance molecules, such as netrin-1. We previously showed that the related netrin-5 (NTN5) is expressed in Mash1-positive transit-amplifying cells and doublecortin-positive neuroblasts in the granule cell layer of the olfactory bulb, the RMS, and the subgranular zone of the adult mouse brain. However, the precise role of NTN5 in adult neurogenesis has not been investigated. In this study, we show that proliferation in the neurogenic niche is impaired in NTN5 knockout mice. The number of proliferating (EdU-labeled) cells in NTN5 KO mice was significantly lower in the V-SVZ, whereas the number of Ki67-positive proliferating cells was unchanged, suggesting a longer cell cycle and decreased cell division in NTN5 KO mice. The number of EdU-labeled cells in the RMS and olfactory bulb was unchanged. By contrast, the numbers of EdU-labeled cells in the cortex, basal ganglia/lateral septal nucleus, and corpus callosum/anterior commissure were increased, which largely represented oligodendrocyte lineage cells. Lastly, we found that chain migration in the RMS of NTN5 KO mice was disorganized. These findings suggest that NTN5 may play important roles in promoting proliferation in the V-SVZ niche, organizing proper chain migration in the RMS, and suppressing oligodendrogenesis in the brain.

    DOI: 10.3389/fnins.2020.570974

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  • Generation of Thyroid Tissues From Embryonic Stem Cells via Blastocyst Complementation In Vivo. 国際誌

    Qingsong Ran, Qiliang Zhou, Kanako Oda, Akihiro Yasue, Manabu Abe, Xulu Ye, Yingchun Li, Toshikuni Sasaoka, Kenji Sakimura, Yoichi Ajioka, Yasuo Saijo

    Frontiers in endocrinology   11   609697 - 609697   2020年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    The generation of mature, functional, thyroid follicular cells from pluripotent stem cells would potentially provide a therapeutic benefit for patients with hypothyroidism, but in vitro differentiation remains difficult. We earlier reported the in vivo generation of lung organs via blastocyst complementation in fibroblast growth factor 10 (Fgf10), compound, heterozygous mutant (Fgf10 Ex1mut/Ex3mut) mice. Fgf10 also plays an essential role in thyroid development and branching morphogenesis, but any role thereof in thyroid organogenesis remains unclear. Here, we report that the thyroids of Fgf10 Ex1mut/Ex3mut mice exhibit severe hypoplasia, and we generate thyroid tissues from mouse embryonic stem cells (ESCs) in Fgf10 Ex1mut/Ex3mut mice via blastocyst complementation. The tissues were morphologically normal and physiologically functional. The thyroid follicular cells of Fgf10 Ex1mut/Ex3mut chimeric mice were derived largely from GFP-positive mouse ESCs although the recipient cells were mixed. Thyroid generation in vivo via blastocyst complementation will aid functional thyroid regeneration.

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  • ジーンターゲッティング法によるGATA4-floxマウスの作製

    岩崎 亜美, 村田 康輔, 川村 名子, 中務 胞, 阿部 学, 夏目 里恵, 杉村 智史, 﨑村 建司, 山城 秀昭

    日本繁殖生物学会 講演要旨集   113 ( 0 )   P - 11-P-11   2020年

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    記述言語:日本語   出版者・発行元:日本繁殖生物学会  

    <p>【目的】GATA4は性腺の分化に関与する転写因子で,生殖巣の発達における重要な役割を担っていると考えられている。一方,胚発生および胎生期の心筋の分化に関与する遺伝子調節も行うため,欠失が起こると,胎生致死となりGATA4ホモ欠損マウスを作出することは極めて困難である。本研究では,卵巣特異的GATA4ホモ欠損マウスの作製のため,ジーンターゲッティング法を用いて,GATA4-floxマウスの作製を行った。このマウスは,Cre-loxpシステムにより,生殖巣特異的にGATA4遺伝子を欠失することが可能となり,胎生致死を回避することができる。【材料・方法】C57/BL6ゲノムライブラリーよりBACクローンを単離,DNAを抽出し,Red/ET組み換え法によるBACサブクローニング法と多重遺伝子導入法を用いてターゲッティングベクターを作製した。エレクトロポレーション法によりES細胞へターゲッティングベクターを導入し,薬剤耐性ES細胞を選抜した。サザンブロット解析により同定した組換えES細胞をICRマウス8細胞期胚に注入し,キメラ胚移植を施した。【結果・考察】その結果,3匹の雄キメラマウスを得た。ES細胞寄与率は,それぞれ約50%,70%,100%であった。自然交配によりキメラマウスとB6雌マウスを交配し,生殖系列キメラマウスを同定し,F1ヘテロマウスの作製に成功した。現在,GATA4-floxヘテロマウス同士を交配させ,ホモマウスを作製している。今後は,生殖巣特異的Vasa-Cre Tg雄マウスとの交配を行い,GATA4欠失マウスの妊孕性の有無および,卵巣形成について組織学的解析を行う予定である。</p>

    DOI: 10.14882/jrds.113.0_P-11

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  • Detection of a High-Turnover Serotonin Circuit in the Mouse Brain Using Mass Spectrometry Imaging. 査読 国際誌

    Eiji Sugiyama, Matteo M Guerrini, Kurara Honda, Yuko Hattori, Manabu Abe, Patrik Källback, Per E Andrén, Kenji F Tanaka, Mitsutoshi Setou, Sidonia Fagarasan, Makoto Suematsu, Yuki Sugiura

    iScience   20   359 - 372   2019年10月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Monoamine neurotransmitters are released by specialized neurons regulating behavioral, motor, and cognitive functions. Although the localization of monoaminergic neurons in the brain is well known, the distribution and kinetics of monoamines remain unclear. Here, we generated a murine brain atlas of serotonin (5-HT), dopamine (DA), and norepinephrine (NE) levels using mass spectrometry imaging (MSI). We found several nuclei rich in both 5-HT and a catecholamine (DA or NE) and identified the paraventricular nucleus of the thalamus (PVT), where 5-HT and NE are co-localized. The analysis of 5-HT fluctuations in response to acute tryptophan depletion and infusion of isotope-labeled tryptophan in vivo revealed a close kinetic association between the raphe nuclei, PVT, and amygdala but not the other nuclei. Our findings imply the existence of a highly dynamic 5-HT-mediated raphe to PVT pathway that likely plays a role in the brain monoamine system.

    DOI: 10.1016/j.isci.2019.09.036

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  • Differential Roles of Each Orexin Receptor Signaling in Obesity. 査読 国際誌

    Miyo Kakizaki, Yousuke Tsuneoka, Kenkichi Takase, Staci J Kim, Jinhwan Choi, Aya Ikkyu, Manabu Abe, Kenji Sakimura, Masashi Yanagisawa, Hiromasa Funato

    iScience   20   1 - 13   2019年10月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Orexins are hypothalamic neuropeptides that regulate feeding, energy expenditure, and sleep. Although orexin-deficient mice are susceptible to obesity, little is known about the roles of the orexin receptors in long-term energy metabolism. Here, we performed the metabolic characterization of orexin receptor-deficient mice. Ox1r-deficient mice were resistant to diet-induced obesity, and their food intake was similar between chow and high-fat food. Ox2r-deficient mice exhibited less energy expenditure than wild-type mice when fed a high-fat diet. Neither Ox1r-deficient nor Ox2r-deficient mice showed body weight gain similar to orexin-deficient mice. Although the presence of a running wheel suppressed diet-induced obesity in wild-type mice, the effect was weaker in orexin neuron-ablated mice. Finally, we did not detect abnormalities in brown adipose tissues of orexin-deficient mice. Thus, each orexin receptor signaling has a unique role in energy metabolism, and orexin neurons are involved in the interactive effect of diet and exercise on body weight gain.

    DOI: 10.1016/j.isci.2019.09.003

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  • Mice deficient in the C-terminal domain of TAR DNA-binding protein 43 develop age-dependent motor dysfunction associated with impaired Notch1-Akt signaling pathway. 査読 国際誌

    Kohei Nishino, Seiji Watanabe, Jin Shijie, Yuri Murata, Kotaro Oiwa, Okiru Komine, Fumito Endo, Hitomi Tsuiji, Manabu Abe, Kenji Sakimura, Amit Mishra, Koji Yamanaka

    Acta neuropathologica communications   7 ( 1 )   118 - 118   2019年7月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Intracellular mislocalization of TAR DNA-binding protein 43 (TDP-43), a nuclear DNA/RNA-binding protein involved in RNA metabolism, is a pathological hallmark of amyotrophic lateral sclerosis (ALS). Although the aggregation-prone, TDP-43 C-terminal domain is widely considered as a key component of TDP-43 pathology in ALS, recent studies including ours suggest that TDP-43 N-terminal fragments (TDP-∆C) may also contribute to the motor dysfunction in ALS. However, the specific pathological functions of TDP-43 N-terminal fragments in mice have not been elucidated. Here, we established TDP-∆C knock-in mice missing a part of exon 6 of murine Tardbp gene, which encodes the C-terminal region of TDP-43. Homozygous TDP-∆C mice showed embryonic lethality, indicating that the N-terminal domain of TDP-43 alone is not sufficient for normal development. In contrast, heterozygous TDP-∆C mice developed normally but exhibited age-dependent mild motor dysfunction with a loss of C-boutons, large cholinergic synaptic terminals on spinal α-motor neurons. TDP-∆C protein broadly perturbed gene expression in the spinal cords of aged heterozygous TDP-∆C mice, including downregulation of Notch1 mRNA. Moreover, the level of Notch1 mRNA was suppressed both by TDP-43 depletion and TDP-∆C expression in Neuro2a cells. Decreased Notch1 mRNA expression in aged TDP-∆C mice was associated with the age-dependent motor dysfunction and loss of Akt surviving signal. Our findings indicate that the N-terminal region of TDP-43 derived from TDP-∆C induces the age-dependent motor dysfunction associated with impaired Notch1-Akt axis in mice.

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  • Mechanical regulation of oligodendrocyte morphology and maturation by the mechanosensor p130Cas. 査読 国際誌

    Takeshi Shimizu, Yasuyuki Osanai, Kenji F Tanaka, Truc Quynh Thai, Manabu Abe, Rie Natsume, Kenji Sakimura, Kazuhiro Ikenaka

    Journal of neurochemistry   150 ( 2 )   158 - 172   2019年7月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:Journal of Neurochemistry  

    Oligodendrocytes (OLs) are myelinating cells of the central nervous system. Recent studies have shown that mechanical factors influence various cell properties. Mechanical stimulation can be transduced into intracellular biochemical signals through mechanosensors, such as integrin, p130Cas, talin and vinculin. However, the molecular mechanisms underlying the mechanical regulation of OLs by mechanosensors remain largely unknown. We found that morphology of OL was affected by knockdown of the mechanosensors p130Cas or talin1. Stretching of OL precursor cells induced the phosphorylation of p130Cas and talin-associated assembly of vinculin. Shear stress decreased the number of OL processes, whereas these effects were mechanically suppressed by dominant-negative (DN) p130Cas, but not by DN-talin1. To investigate the roles of p130Cas in post-natal OLs in vivo, we constructed a novel p130Cas knock-in mouse and found overexpression of p130Cas in vivo affected the number of mature OLs in the cortex. These results indicate that the mechanosensor p130Cas controls both OL morphogenesis and maturation.

    DOI: 10.1111/jnc.14657

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  • Dissociating orexin-dependent and -independent functions of orexin neurons using novel Orexin-Flp knock-in mice. 査読 国際誌

    Srikanta Chowdhury, Chi Jung Hung, Shuntaro Izawa, Ayumu Inutsuka, Meiko Kawamura, Takashi Kawashima, Haruhiko Bito, Itaru Imayoshi, Manabu Abe, Kenji Sakimura, Akihiro Yamanaka

    eLife   8   2019年6月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Uninterrupted arousal is important for survival during threatening situations. Activation of orexin/hypocretin neurons is implicated in sustained arousal. However, orexin neurons produce and release orexin as well as several co-transmitters including dynorphin and glutamate. To disambiguate orexin-dependent and -independent physiological functions of orexin neurons, we generated a novel Orexin-flippase (Flp) knock-in mouse line. Crossing with Flp-reporter or Cre-expressing mice showed gene expression exclusively in orexin neurons. Histological studies confirmed that orexin was knock-out in homozygous mice. Orexin neurons without orexin showed altered electrophysiological properties, as well as received decreased glutamatergic inputs. Selective chemogenetic activation revealed that both orexin and co-transmitters functioned to increase wakefulness, however, orexin was indispensable to promote sustained arousal. Surprisingly, such activation increased the total time spent in cataplexy. Taken together, orexin is essential to maintain basic membrane properties and input-output computation of orexin neurons, as well as to exert awake-sustaining aptitude of orexin neurons.

    DOI: 10.7554/eLife.44927

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  • GABA neurons in the ventral tegmental area regulate non-rapid eye movement sleep in mice. 査読 国際誌

    Srikanta Chowdhury, Takanori Matsubara, Toh Miyazaki, Daisuke Ono, Noriaki Fukatsu, Manabu Abe, Kenji Sakimura, Yuki Sudo, Akihiro Yamanaka

    eLife   8   2019年6月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Sleep/wakefulness cycle is regulated by coordinated interactions between sleep- and wakefulness-regulating neural circuitry. However, the detailed mechanism is far from understood. Here, we found that glutamic acid decarboxylase 67-positive GABAergic neurons in the ventral tegmental area (VTAGad67+) are a key regulator of non-rapid eye movement (NREM) sleep in mice. VTAGad67+ project to multiple brain areas implicated in sleep/wakefulness regulation such as the lateral hypothalamus (LH). Chemogenetic activation of VTAGad67+ promoted NREM sleep with higher delta power whereas optogenetic inhibition of these induced prompt arousal from NREM sleep, even under highly somnolescent conditions, but not from REM sleep. VTAGad67+ showed the highest activity in NREM sleep and the lowest activity in REM sleep. Moreover, VTAGad67+ directly innervated and inhibited wake-promoting orexin/hypocretin neurons by releasing GABA. As such, optogenetic activation of VTAGad67+ terminals in the LH promoted NREM sleep. Taken together, we revealed that VTAGad67+ play an important role in the regulation of NREM sleep.

    DOI: 10.7554/eLife.44928

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  • Perturbed expression pattern of the immediate early gene Arc in the dentate gyrus of GluA1 C-terminal palmitoylation-deficient mice. 査読 国際誌

    Masayuki Itoh, Hiroyuki Okuno, Daisuke Yamada, Mariko Yamashita, Manabu Abe, Rie Natsume, Toshie Kaizuka, Kenji Sakimura, Mikio Hoshino, Masayoshi Mishina, Keiji Wada, Masayuki Sekiguchi, Takashi Hayashi

    Neuropsychopharmacology reports   39 ( 1 )   61 - 66   2019年3月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    BACKGROUND: AMPA receptors predominantly mediate fast excitatory synaptic transmission in the mammalian brain. Post-translational protein S-palmitoylation of AMPA receptor GluA subunits at their C-termini reversibly controls the receptors trafficking to and from excitatory glutamatergic synapses. Excitatory inputs to neurons induce the expression of immediate early genes (IEGs), including Arc, with particular spatial patterns. In the hippocampal dentate gyrus, Arc is mainly expressed in the upper (dorsal) blade at the basal state. GluA1 C-terminal palmitoylation-deficient (GluA1C811S) mice showed enhanced seizure susceptibility and disturbed synaptic plasticity without impaired gross anatomy or basal synaptic transmission. These mutant mice also exhibited an increased expression of IEG products, c-Fos and Arc proteins, in the hippocampus and cerebral cortex. In this report, we further analyzed excitability and Arc expression pattern in the dentate gyrus of GluA1C811S mice. METHODS AND RESULTS: Electrophysiological analysis of granule neurons to measure the evoked excitatory postsynaptic current/evoked inhibitory postsynaptic current ratio revealed that excitatory/inhibitory (E/I) balance was normal in GluA1C811S mice. In contrast, immunohistochemical staining showed an abnormal distribution of Arc-positive cells between upper and lower (ventral) blades of the dentate gyrus in these mutant mice. These data suggest that deficiency of GluA1 palmitoylation causes perturbed neuronal inputs from the entorhinal cortex to the dentate gyrus, which potentially underlies the excessive excitability in response to seizure-inducing stimulation. CONCLUSION: Our findings conclude that an appropriate regulation of Arc expression in the dentate gyrus, ensured by AMPA receptor palmitoylation, may be critical for stabilizing hippocampal neural circuits and may suppress excess excitation.

    DOI: 10.1002/npr2.12044

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  • Cathepsin C modulates myelin oligodendrocyte glycoprotein-induced experimental autoimmune encephalomyelitis. 査読 国際誌

    Wilaiwan Wisessmith Durose, Takahiro Shimizu, JiaYi Li, Manabu Abe, Kenji Sakimura, Banthit Chetsawang, Kenji F Tanaka, Akio Suzumura, Koujiro Tohyama, Kazuhiro Ikenaka

    Journal of neurochemistry   148 ( 3 )   413 - 425   2019年2月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Multiple sclerosis (MS) is an autoimmune disease characterized by immune-mediated inflammation, which attacks the myelin sheath. MS pursues a relapsing and remitting course with varying intervals between symptoms. The main clinical pathological features include inflammation, myelin sheath destruction and plaque formation in the central nervous system (CNS). We previously reported that cystatin F (CysF) expression is induced in demyelinating lesions that are accompanied by active remyelination (referred to as shadow plaques) but is down-regulated in chronic demyelinated lesions (plaques) in the spinal cord of MS patients and in several murine models of demyelinating disease. CysF is a cathepsin protease inhibitor whose major target is cathepsin C (CatC), which is co-expressed in demyelinating regions in Plp4e/- mice, a model of chronic demyelination. Here, we report the time course of CatC and CysF expression and describe the symptoms in a mouse experimental autoimmune encephalomyelitis (EAE) model using CatC knockdown (KD) and CatC over-expression (OE) mice. In myelin oligodendrocyte glycoprotein (MOG)-EAE, CatC positive cells were found to infiltrate the CNS at an early stage prior to any clinical signs, in comparison to WT mice. CysF expression was not observed at this early stage, but appeared later within shadow plaques. CatC expression was found in chronic demyelinated lesions but was not associated with CysF expression, and CatCKD EAE mouse showed delayed demyelination. Whereas, CatCOE in microglia significantly increased severity of demyelination in the MOG-EAE model. Thus, these results demonstrate that CatC plays a major role in MOG-EAE.

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  • ELKS/Voltage-Dependent Ca2+ Channel-β Subunit Module Regulates Polarized Ca2+ Influx in Pancreatic β Cells. 査読 国際誌

    Mica Ohara-Imaizumi, Kyota Aoyagi, Hajime Yamauchi, Masashi Yoshida, Masayuki X Mori, Yamato Hida, Ha Nam Tran, Masamichi Ohkura, Manabu Abe, Yoshihiro Akimoto, Yoko Nakamichi, Chiyono Nishiwaki, Hayato Kawakami, Kazuo Hara, Kenji Sakimura, Shinya Nagamatsu, Yasuo Mori, Junichi Nakai, Masafumi Kakei, Toshihisa Ohtsuka

    Cell reports   26 ( 5 )   1213 - 1226   2019年1月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:CELL PRESS  

    Pancreatic β cells secrete insulin by Ca2+-triggered exocytosis. However, there is no apparent secretory site similar to the neuronal active zones, and the cellular and molecular localization mechanism underlying polarized exocytosis remains elusive. Here, we report that ELKS, a vertebrate active zone protein, is used in β cells to regulate Ca2+ influx for insulin secretion. β cell-specific ELKS-knockout (KO) mice showed impaired glucose-stimulated first-phase insulin secretion and reduced L-type voltage-dependent Ca2+ channel (VDCC) current density. In situ Ca2+ imaging of β cells within islets expressing a membrane-bound G-CaMP8b Ca2+ sensor demonstrated initial local Ca2+ signals at the ELKS-localized vascular side of the β cell plasma membrane, which were markedly decreased in ELKS-KO β cells. Mechanistically, ELKS directly interacted with the VDCC-β subunit via the GK domain. These findings suggest that ELKS and VDCCs form a potent insulin secretion complex at the vascular side of the β cell plasma membrane for polarized Ca2+ influx and first-phase insulin secretion from pancreatic islets.

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  • Autophosphorylation of F-actin binding domain of CaMKIIβ is required for fear learning. 査読 国際誌

    Kim K, Suzuki A, Kojima H, Kawamura M, Miya K, Abe M, Yamada I, Furuse T, Wakana S, Sakimura K, Hayashi Y

    Neurobiology of learning and memory   157   86 - 95   2019年1月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    DOI: 10.1016/j.nlm.2018.12.003

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  • The role of sphingosine-1-phosphate in inflammation and cancer progression. 査読 国際誌

    Masayuki Nagahashi, Manabu Abe, Kenji Sakimura, Kazuaki Takabe, Toshifumi Wakai

    Cancer science   109 ( 12 )   3671 - 3678   2018年12月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Many inflammatory mediators are involved in the process of carcinogenesis and cancer progression. In addition to cytokines and chemokines, lipid mediators have recently attracted attention as signaling molecules associated with inflammatory diseases. Sphingosine-1-phosphate (S1P) is a pleiotropic lipid mediator that regulates cell survival and migration, immune cell recruitment, angiogenesis and lymphangiogenesis. S1P also plays a significant role in inflammation and cancer. The gradation of S1P concentration in the blood, lymph and tissue regulates lymphocyte trafficking, an important component of inflammation. Furthermore, cancer cells produce elevated levels of S1P, contributing to the tumor microenvironment and linking cancer and inflammation. Future technological advances may reveal greater detail about the mechanisms of S1P regulation in the tumor microenvironment and the contribution of S1P to cancer progression. Considering the critical role of S1P in linking inflammation and cancer, it is possible that the S1P signaling pathway could be a novel therapeutic target for cancers with chronic inflammation.

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  • Different Roles of Sphingosine Kinase 1 and 2 in Pancreatic Cancer Progression. 査読 国際誌

    Kizuki Yuza, Masato Nakajima, Masayuki Nagahashi, Junko Tsuchida, Yuki Hirose, Kohei Miura, Yosuke Tajima, Manabu Abe, Kenji Sakimura, Kazuaki Takabe, Toshifumi Wakai

    The Journal of surgical research   232   186 - 194   2018年12月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    BACKGROUND: Pancreatic cancer is a disease with poor prognosis, and development of new treatments is necessary. Sphingosine-1-phosphate (S1P), a bioactive lipid mediator produced by sphingosine kinases (SphK1 and SphK2), plays a critical role in progression of many types of cancer. However, little is known about the role of sphingosine kinases in pancreatic cancer. This study investigated the roles of sphingosine kinases in pancreatic cancer progression. MATERIALS AND METHODS: S1P levels in pancreatic cancer and noncancerous pancreatic tissue were measured in 10 patients. We generated PAN02 murine pancreatic cancer cell lines with a clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated system genes 9 (Cas9)-mediated deletion of SphK1 or SphK2 and assessed cell growth and migration. In an animal model, we assessed the survival of mice injected with PAN02 cells intraperitoneally. RESULTS: S1P levels in the pancreatic cancer tissue were significantly higher than those in noncancerous tissue. SphK1 knockout (KO) cells showed greater proliferation and migration than wild type (WT) cells, and SphK2 KO cells showed less proliferation and migration than WT cells. Animal experiments showed that the survival of mice injected with SphK1 KO cells was significantly shorter than those injected with WT cells, and the survival of mice injected with SphK2 KO cells was longer than those injected with WT cells. Surprisingly, cytotoxic assay using gemcitabine showed that SphK1 KO cells survived less than WT cells, and SphK2 KO cells survived more than WT cells. CONCLUSIONS: S1P produced by SphK1 and SphK2 may have different functions in pancreatic cancer cells. Targeting both SphK1 and SphK2 may be a potential strategy for pancreatic cancer treatment.

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  • Deficiency of AMPAR-Palmitoylation Aggravates Seizure Susceptibility. 査読 国際誌

    Masayuki Itoh, Mariko Yamashita, Masaki Kaneko, Hiroyuki Okuno, Manabu Abe, Maya Yamazaki, Rie Natsume, Daisuke Yamada, Toshie Kaizuka, Reiko Suwa, Kenji Sakimura, Masayuki Sekiguchi, Keiji Wada, Mikio Hoshino, Masayoshi Mishina, Takashi Hayashi

    The Journal of neuroscience : the official journal of the Society for Neuroscience   38 ( 47 )   10220 - 10235   2018年11月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Synaptic AMPAR expression controls the strength of excitatory synaptic transmission and plasticity. An excess of synaptic AMPARs leads to epilepsy in response to seizure-inducible stimulation. The appropriate regulation of AMPARs plays a crucial role in the maintenance of the excitatory/inhibitory synaptic balance; however, the detailed mechanisms underlying epilepsy remain unclear. Our previous studies have revealed that a key modification of AMPAR trafficking to and from postsynaptic membranes is the reversible, posttranslational S-palmitoylation at the C-termini of receptors. To clarify the role of palmitoylation-dependent regulation of AMPARs in vivo, we generated GluA1 palmitoylation-deficient (Cys811 to Ser substitution) knock-in mice. These mutant male mice showed elevated seizure susceptibility and seizure-induced neuronal activity without impairments in synaptic transmission, gross brain structure, or behavior at the basal level. Disruption of the palmitoylation site was accompanied by upregulated GluA1 phosphorylation at Ser831, but not at Ser845, in the hippocampus and increased GluA1 protein expression in the cortex. Furthermore, GluA1 palmitoylation suppressed excessive spine enlargement above a certain size after LTP. Our findings indicate that an abnormality in GluA1 palmitoylation can lead to hyperexcitability in the cerebrum, which negatively affects the maintenance of network stability, resulting in epileptic seizures.SIGNIFICANCE STATEMENT AMPARs predominantly mediate excitatory synaptic transmission. AMPARs are regulated in a posttranslational, palmitoylation-dependent manner in excitatory synapses of the mammalian brain. Reversible palmitoylation dynamically controls synaptic expression and intracellular trafficking of the receptors. Here, we generated GluA1 palmitoylation-deficient knock-in mice to clarify the role of AMPAR palmitoylation in vivo We showed that an abnormality in GluA1 palmitoylation led to hyperexcitability, resulting in epileptic seizure. This is the first identification of a specific palmitoylated protein critical for the seizure-suppressing process. Our data also provide insight into how predicted receptors such as AMPARs can effectively preserve network stability in the brain. Furthermore, these findings help to define novel key targets for developing anti-epileptic drugs.

    DOI: 10.1523/JNEUROSCI.1590-18.2018

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  • Cytomatrix proteins CAST and ELKS regulate retinal photoreceptor development and maintenance. 査読 国際誌

    Akari Hagiwara, Yosuke Kitahara, Chad Paul Grabner, Christian Vogl, Manabu Abe, Ryo Kitta, Keisuke Ohta, Keiichiro Nakamura, Kenji Sakimura, Tobias Moser, Akinori Nishi, Toshihisa Ohtsuka

    The Journal of cell biology   217 ( 11 )   3993 - 4006   2018年11月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    At the presynaptic active zone (AZ), the related cytomatrix proteins CAST and ELKS organize the presynaptic release machinery. While CAST is known to regulate AZ size and neurotransmitter release, the role of ELKS and the integral system of CAST/ELKS together is poorly understood. Here, we show that CAST and ELKS have both redundant and unique roles in coordinating synaptic development, function, and maintenance of retinal photoreceptor ribbon synapses. A CAST/ELKS double knockout (dKO) mouse showed high levels of ectopic synapses and reduced responses to visual stimulation. Ectopic formation was not observed in ELKS conditional KO but progressively increased with age in CAST KO mice with higher rates in the dKO. Presynaptic calcium influx was strongly reduced in rod photoreceptors of CAST KO and dKO mice. Three-dimensional scanning EM reconstructions showed structural abnormalities in rod triads of CAST KO and dKO. Remarkably, AAV-mediated acute ELKS deletion after synapse maturation induced neurodegeneration and loss of ribbon synapses. These results suggest that CAST and ELKS work in concert to promote retinal synapse formation, transmission, and maintenance.

    DOI: 10.1083/jcb.201704076

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  • Deletion of exons encoding carboxypeptidase domain of Nna1 results in Purkinje cell degeneration (pcd) phenotype. 査読 国際誌

    Li Zhou, M Ibrahim Hossain, Maya Yamazaki, Manabu Abe, Rie Natsume, Kohtaro Konno, Shun Kageyama, Masaaki Komatsu, Masahiko Watanabe, Kenji Sakimura, Hirohide Takebayashi

    Journal of neurochemistry   147 ( 4 )   557 - 572   2018年11月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Purkinje cell degeneration (pcd) was first identified in a spontaneous mouse mutant showing cerebellar ataxia. In addition to cerebellar Purkinje cells (PCs), retinal photoreceptors, mitral cells in the olfactory bulb, and a discrete subpopulation of thalamic neurons also degenerate in the mutant brains. The gene responsible for the pcd mutant is Nna1, also known as ATP/GTP binding protein 1 or cytosolic carboxypeptidase-like 1, which encodes a zinc carboxypeptidase protein. To investigate pathogenesis of the pcd mutation in detail, we generated a conditional Nna1 allele targeting the carboxypeptidase domain at C-terminus. After Cre recombination and heterozygous crossing, we generated Nna1 knockout (KO) mice and found that the Nna1 KO mice began to show cerebellar ataxia at postnatal day 20 (P20). Most PCs degenerated until 4-week-old, except lobule X. Activated microglia and astrocytes were also observed in the Nna1 KO cerebellum. In the mutant brain, the Nna1 mRNA level was dramatically reduced, suggesting that nonsense-mediated mRNA decay occurs in it. Since the Nna1 protein acts as a de-glutamatase on the C-terminus of α-tubulin and β-tubulin, increased polyglutamylated tubulin was detected in the Nna1 KO cerebellum. In addition, the endoplasmic reticulum stress marker, C/EBP homologous protein (CHOP), was up-regulated in the mutant PCs. We report the generation of a functional Nna1 conditional allele and possible mechanisms of PC death in the Nna1 KO in the cerebellum. OPEN PRACTICES: This article has received a badge for *Open Materials* because it provided all relevant information to reproduce the study in the manuscript. The complete Open Science Disclosure form for this article can be found at the end of the article. More information about the Open Practices badges can be found at https://cos.io/our-services/open-science-badges/.

    DOI: 10.1111/jnc.14591

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  • Distribution of Caskin1 protein and phenotypic characterization of its knockout mice using a comprehensive behavioral test battery. 査読 国際誌

    Tayo Katano, Keizo Takao, Manabu Abe, Maya Yamazaki, Masahiko Watanabe, Tsuyoshi Miyakawa, Kenji Sakimura, Seiji Ito

    Molecular brain   11 ( 1 )   63 - 63   2018年10月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Calcium/calmodulin-dependent serine protein kinase (CASK)-interacting protein 1 (Caskin1) is a direct binding partner of the synaptic adaptor protein CASK. Because Caskin1 forms homo-multimers and binds not only CASK but also other neuronal proteins in vitro, it is anticipated to have neural functions; but its exact role in mammals remains unclear. Previously, we showed that the concentration of Caskin1 in the spinal dorsal horn increases under chronic pain. To characterize this protein, we generated Caskin1-knockout (Caskin1-KO) mice and specific anti-Caskin1 antibodies. Biochemical and immunohistochemical analyses demonstrated that Caskin1 was broadly distributed in the whole brain and spinal cord, and that it primarily localized at synapses. To elucidate the neural function of Caskin1 in vivo, we subjected Caskin1-KO mice to comprehensive behavioral analysis. The mutant mice exhibited differences in gait, enhanced nociception, and anxiety-like behavior relative to their wild-type littermates. In addition, the knockouts exhibited strong freezing responses, with or without a cue tone, in contextual and cued-fear conditioning tests as well as low memory retention in the Barnes Maze test. Taken together, these results suggest that Caskin1 contributes to a wide spectrum of behavioral phenotypes, including gait, nociception, memory, and stress response, in broad regions of the central nervous system.

    DOI: 10.1186/s13041-018-0407-2

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  • CDC42EP4, a perisynaptic scaffold protein in Bergmann glia, is required for glutamatergic tripartite synapse configuration. 査読 国際誌

    Natsumi Ageta-Ishihara, Kohtarou Konno, Maya Yamazaki, Manabu Abe, Kenji Sakimura, Masahiko Watanabe, Makoto Kinoshita

    Neurochemistry international   119   190 - 198   2018年10月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Configuration of tripartite synapses, comprising the pre-, post-, and peri-synaptic components (axon terminal or bouton, dendritic spine, and astroglial terminal process), is a critical determinant of neurotransmitter kinetics and hence synaptic transmission. However, little is known about molecular basis for the regulation of tripartite synapse morphology. Previous studies showed that CDC42EP4, an effector protein of a cell morphogenesis regulator CDC42, is expressed exclusively in Bergmann glia in the cerebellar cortex, that it forms tight complex with the septin heterooligomer, and that it interacts indirectly with the glutamate transporter GLAST and MYH10/nonmuscle myosin ΙΙB. Scrutiny of Cdc42ep4-/- mice had revealed that the CDC42EP4-septins-GLAST interaction facilitates glutamate clearance, while the role for CDC42EP4-septins-MYH10 interaction has remained unsolved. Here, we find anomalous configuration of the tripartite synapses comprising the parallel fiber boutons, dendritic spines of Purkinje cells, and Bergmann glial processes in Cdc42ep4-/- mice. The complex anomalies include 1) recession of Bergmann glial membranes from the nearest active zones, and 2) extension of nonactive synaptic contact around active zone. In line with the recession of Bergmann glial membranes by the loss of CDC42EP4, overexpression of CDC42EP4 in heterologous cells promotes cell spreading and partitioning of MYH10 to insoluble (i.e., active) fraction. Paradoxically, however, Cdc42ep4-/- cerebellum contained significantly more MYH10 and N-cadherin, which is attributed to secondary neuronal response mainly in Purkinje cells. Given cooperative actions of N-cadherin and MYH10 for adhesion between neurons, we speculate that their augmentation may reflect the extension of nonactive synaptic contacts in Cdc42ep4-/- cerebellum. Transcellular mechanism that links the absence of CDC42EP4 in Bergmann glia to the augmentation of N-cadherin and MYH10 in neurons is currently unknown, but the phenotypic similarity to GLAST-null mice indicates involvement of the glutamate intolerance. Together, the unique phenotype of Cdc42ep4-/- mice provides a clue to novel molecular network underlying tripartite synapse configuration.

    DOI: 10.1016/j.neuint.2018.01.003

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  • Ablation of Central Serotonergic Neurons Decreased REM Sleep and Attenuated Arousal Response

    Iwasaki, Kanako, Komiya, Haruna, Kakizaki, Miyo, Miyoshi, Chika, Abe, Manabu, Sakimura, Kenji, Funato, Hiromasa, Yanagisawa, Masashi

    FRONTIERS IN NEUROSCIENCE   12   2018年8月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:FRONTIERS MEDIA SA  

    DOI: 10.3389/fnins.2018.00535

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  • Higher visual responses in the temporal cortex of mice. 査読 国際誌

    Nana Nishio, Hiroaki Tsukano, Ryuichi Hishida, Manabu Abe, Junichi Nakai, Meiko Kawamura, Atsushi Aiba, Kenji Sakimura, Katsuei Shibuki

    Scientific reports   8 ( 1 )   11136 - 11136   2018年7月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    The visual cortex of mice is a useful model for investigating the mammalian visual system. In primates, higher visual areas are classified into two parts, the dorsal stream ("where" pathway) and ventral stream ("what" pathway). The ventral stream is known to include a part of the temporal cortex. In mice, however, some cortical areas adjacent to the primary visual area (V1) in the occipital cortex are thought to be comparable to the ventral stream in primates, although the whole picture of the mouse ventral stream has never been elucidated. We performed wide-field Ca2+ imaging in awake mice to investigate visual responses in the mouse temporal cortex, and found that the postrhinal cortex (POR), posterior to the auditory cortex (AC), and the ectorhinal and temporal association cortices (ECT), ventral to the AC, showed clear visual responses to moving visual objects. The retinotopic maps in the POR and ECT were not clearly observed, and the amplitudes of the visual responses in the POR and ECT were less sensitive to the size of the objects, compared to visual responses in the V1. In the ECT, objects of different sizes activated different subareas. These findings strongly suggest that the mouse ventral stream extends to the ECT ventral to the AC, and that it has characteristic response properties that are markedly different from the response properties in the V1.

    DOI: 10.1038/s41598-018-29530-3

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  • CAST/ELKS Proteins Control Voltage-Gated Ca2+ Channel Density and Synaptic Release Probability at a Mammalian Central Synapse. 査読 国際誌

    Wei Dong, Tamara Radulovic, R Oliver Goral, Connon Thomas, Monica Suarez Montesinos, Debbie Guerrero-Given, Akari Hagiwara, Travis Putzke, Yamato Hida, Manabu Abe, Kenji Sakimura, Naomi Kamasawa, Toshihisa Ohtsuka, Samuel M Young Jr

    Cell reports   24 ( 2 )   284 - 293   2018年7月

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    記述言語:英語  

    In the presynaptic terminal, the magnitude and location of Ca2+ entry through voltage-gated Ca2+ channels (VGCCs) regulate the efficacy of neurotransmitter release. However, how presynaptic active zone proteins control mammalian VGCC levels and organization is unclear. To address this, we deleted the CAST/ELKS protein family at the calyx of Held, a CaV2.1 channel-exclusive presynaptic terminal. We found that loss of CAST/ELKS reduces the CaV2.1 current density with concomitant reductions in CaV2.1 channel numbers and clusters. Surprisingly, deletion of CAST/ELKS increases release probability while decreasing the readily releasable pool, with no change in active zone ultrastructure. In addition, Ca2+ channel coupling is unchanged, but spontaneous release rates are elevated. Thus, our data identify distinct roles for CAST/ELKS as positive regulators of CaV2.1 channel density and suggest that they regulate release probability through a post-priming step that controls synaptic vesicle fusogenicity.

    DOI: 10.1016/j.celrep.2018.06.024

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  • Targeting the SphK1/S1P/S1PR1 Axis That Links Obesity, Chronic Inflammation, and Breast Cancer Metastasis. 査読 国際誌

    Masayuki Nagahashi, Akimitsu Yamada, Eriko Katsuta, Tomoyoshi Aoyagi, Wei-Ching Huang, Krista P Terracina, Nitai C Hait, Jeremy C Allegood, Junko Tsuchida, Kizuki Yuza, Masato Nakajima, Manabu Abe, Kenji Sakimura, Sheldon Milstien, Toshifumi Wakai, Sarah Spiegel, Kazuaki Takabe

    Cancer research   78 ( 7 )   1713 - 1725   2018年4月

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    記述言語:英語  

    Although obesity with associated inflammation is now recognized as a risk factor for breast cancer and distant metastases, the functional basis for these connections remain poorly understood. Here, we show that in breast cancer patients and in animal breast cancer models, obesity is a sufficient cause for increased expression of the bioactive sphingolipid mediator sphingosine-1-phosphate (S1P), which mediates cancer pathogenesis. A high-fat diet was sufficient to upregulate expression of sphingosine kinase 1 (SphK1), the enzyme that produces S1P, along with its receptor S1PR1 in syngeneic and spontaneous breast tumors. Targeting the SphK1/S1P/S1PR1 axis with FTY720/fingolimod attenuated key proinflammatory cytokines, macrophage infiltration, and tumor progression induced by obesity. S1P produced in the lung premetastatic niche by tumor-induced SphK1 increased macrophage recruitment into the lung and induced IL6 and signaling pathways important for lung metastatic colonization. Conversely, FTY720 suppressed IL6, macrophage infiltration, and S1P-mediated signaling pathways in the lung induced by a high-fat diet, and it dramatically reduced formation of metastatic foci. In tumor-bearing mice, FTY720 similarly reduced obesity-related inflammation, S1P signaling, and pulmonary metastasis, thereby prolonging survival. Taken together, our results establish a critical role for circulating S1P produced by tumors and the SphK1/S1P/S1PR1 axis in obesity-related inflammation, formation of lung metastatic niches, and breast cancer metastasis, with potential implications for prevention and treatment.Significance: These findings offer a preclinical proof of concept that signaling by a sphingolipid may be an effective target to prevent obesity-related breast cancer metastasis. Cancer Res; 78(7); 1713-25. ©2018 AACR.

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  • 胚盤胞補完法を利用したマウスES細胞による肺再生

    北原 哲彦, 周 啓亮, 冉 慶松, 叶 許緑, 小田 佳奈子, 笹岡 俊邦, 阿部 学, 崎村 建司, 味岡 洋一, 泰江 章博, 土田 正則, 西條 康夫

    日本外科学会定期学術集会抄録集   118回   1340 - 1340   2018年4月

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    記述言語:日本語   出版者・発行元:(一社)日本外科学会  

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  • Retrograde Signaling from Progranulin to Sort1 Counteracts Synapse Elimination in the Developing Cerebellum. 査読 国際誌

    Naofumi Uesaka, Manabu Abe, Kohtarou Konno, Maya Yamazaki, Kazuto Sakoori, Takaki Watanabe, Tzu-Huei Kao, Takayasu Mikuni, Masahiko Watanabe, Kenji Sakimura, Masanobu Kano

    Neuron   97 ( 4 )   796 - 805   2018年2月

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    記述言語:英語  

    Elimination of redundant synapses formed early in development and strengthening of necessary connections are crucial for shaping functional neural circuits. Purkinje cells (PCs) in the neonatal cerebellum are innervated by multiple climbing fibers (CFs) with similar strengths. A single CF is strengthened whereas the other CFs are eliminated in each PC during postnatal development. The underlying mechanisms, particularly for the strengthening of single CFs, are poorly understood. Here we report that progranulin, a multi-functional growth factor implicated in the pathogenesis of frontotemporal dementia, strengthens developing CF synaptic inputs and counteracts their elimination from postnatal day 11 to 16. Progranulin derived from PCs acts retrogradely onto its putative receptor Sort1 on CFs. This effect is independent of semaphorin 3A, another retrograde signaling molecule that counteracts CF synapse elimination. We propose that progranulin-Sort1 signaling strengthens and maintains developing CF inputs, and may contribute to selection of single "winner" CFs that survive synapse elimination.

    DOI: 10.1016/j.neuron.2018.01.018

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  • Ablation of Central Serotonergic Neurons Decreased REM Sleep and Attenuated Arousal Response. 査読 国際誌

    Kanako Iwasaki, Haruna Komiya, Miyo Kakizaki, Chika Miyoshi, Manabu Abe, Kenji Sakimura, Hiromasa Funato, Masashi Yanagisawa

    Frontiers in neuroscience   12   535 - 535   2018年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Sleep/wake behavior is regulated by distinct groups of neurons, such as dopaminergic, noradrenergic, and orexinergic neurons. Although monoaminergic neurons are usually considered to be wake-promoting, the role of serotonergic neurons in sleep/wake behavior remains inconclusive because of the effect of serotonin (5-HT)-deficiency on brain development and the compensation for inborn 5-HT deficiency by other sleep/wake-regulating neurons. Here, we performed selective ablation of central 5-HT neurons in the newly developed Rosa-diphtheria toxin receptor (DTR)-tdTomato mouse line that was crossed with Pet1Cre/+ mice to examine the role of 5-HT neurons in the sleep/wake behavior of adult mice. Intracerebroventricular administration of diphtheria toxin completely ablated tdTomato-positive cells in Pet1Cre/+; Rosa-DTR-tdTomato mice. Electroencephalogram/electromyogram-based sleep/wake analysis demonstrated that central 5-HT neuron ablation in adult mice decreased the time spent in rapid eye movement (REM) sleep, which was associated with fewer transitions from non-REM (NREM) sleep to REM sleep than in control mice. Central 5-HT neuron-ablated mice showed attenuated wake response to a novel environment and increased theta power during wakefulness compared to control mice. The current findings indicated that adult 5-HT neurons work to support wakefulness and regulate REM sleep time through a biased transition from NREM sleep to REM sleep.

    DOI: 10.3389/fnins.2018.00535

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  • Serotonergic Input to Orexin Neurons Plays a Role in Maintaining Wakefulness and REM Sleep Architecture. 査読 国際誌

    Yuki C Saito, Natsuko Tsujino, Manabu Abe, Maya Yamazaki, Kenji Sakimura, Takeshi Sakurai

    Frontiers in neuroscience   12   892 - 892   2018年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:FRONTIERS MEDIA SA  

    Neurons expressing neuropeptide orexins (hypocretins) in the lateral hypothalamus (LH) and serotonergic neurons in the dorsal raphe nucleus (DR) both play important roles in the regulation of sleep/wakefulness states, and show similar firing patterns across sleep/wakefulness states. Orexin neurons send excitatory projections to serotonergic neurons in the DR, which express both subtypes of orexin receptors (Mieda et al., 2011), while serotonin (5-HT) potently inhibits orexin neurons through activation of 5HT1A receptors (5HT1ARs). In this study, we examined the physiological importance of serotonergic inhibitory regulation of orexin neurons by studying the phenotypes of mice lacking the 5HT1A receptor gene (Htr1a) specifically in orexin neurons (ox5HT1ARKO mice). ox5HT1ARKO mice exhibited longer NREM sleep time along with decreased wakefulness time in the later phase of the dark period. We also found that restraint stress induced a larger impact on REM sleep architecture in ox5HT1ARKO mice than in controls, with a larger delayed increase in REM sleep amount as compared with that in controls, indicating abnormality of REM sleep homeostasis in the mutants. These results suggest that 5HT1ARs in orexin neurons are essential in the regulation of sleep/wakefulness states, and that serotonergic regulation of orexin neurons plays a crucial role in the appropriate control of orexinergic tone to maintain normal sleep/wake architecture.

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  • Potentiation of excitatory synaptic transmission ameliorates aggression in mice with Stxbp1 haploinsufficiency. 査読 国際誌

    Hiroyuki Miyamoto, Atsushi Shimohata, Manabu Abe, Teruo Abe, Emi Mazaki, Kenji Amano, Toshimitsu Suzuki, Tetsuya Tatsukawa, Shigeyoshi Itohara, Kenji Sakimura, Kazuhiro Yamakawa

    Human molecular genetics   26 ( 24 )   4961 - 4974   2017年12月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:OXFORD UNIV PRESS  

    Genetic studies point to a major role of de novo mutations in neurodevelopmental disorders of intellectual disability, autism spectrum disorders, and epileptic encephalopathy. The STXBP1 gene encodes the syntaxin-binding protein 1 (Munc18-1) that critically controls synaptic vesicle exocytosis and synaptic transmission. This gene harbors a high frequency of de novo mutations, which may play roles in these neurodevelopmental disorders. However, the system and behavioral-level pathophysiological changes caused by these genetic defects remain poorly understood. Constitutional (Stxbp1+/-), dorsal-telencephalic excitatory (Stxbp1fl/+/Emx), or global inhibitory neuron-specific (Stxbp1fl/+/Vgat) mice were subjected to a behavioral test battery examining locomotor activity, anxiety, fear learning, and social interactions including aggression. Furthermore, measurements of local field potentials in multiple regions of the brain were performed. Stxbp1+/- male mice exhibited enhanced aggressiveness and impaired fear learning associated with elevated gamma activity in several regions of the brain including the prefrontal cortex. Stxbp1fl/+/Emx mice showed fear-learning deficits, but neither Stxbp1fl/+/Emx nor Stxbp1fl/+/Vgat mice showed increased aggressiveness. Pharmacological potentiation of the excitatory transmission at active synapses via the systemic administration of ampakine CX516, which enhances the excitatory postsynaptic function, ameliorated the aggressive phenotype of Stxbp1+/- mice. These findings suggest that synaptic impairments of the dorsal telencephalic and subcortical excitatory neurons cause learning deficits and enhanced aggression in Stxbp1+/- mice, respectively. Additionally, normalizing the excitatory synaptic transmission is a potential therapeutic option for managing aggressiveness in patients with STXBP1 mutations.

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  • D1/D2ドーパミン受容体コンディショナル発現マウスによる運動制御機構の解明

    笹岡 俊邦, 佐藤 朝子, 知見 聡美, 大久保 直, 阿部 学, 川村 名子, 中尾 聡宏, 齊藤 奈英, 酒井 清子, 小田 佳奈子, 前田 宜俊, 神保 幸弘, 田中 稔, 山本 美丘, 佐藤 俊哉, 藤澤 信義, 崎村 建司, 南部 篤

    生命科学系学会合同年次大会   2017年度   [4LT08 - 1195)]   2017年12月

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    記述言語:英語   出版者・発行元:生命科学系学会合同年次大会運営事務局  

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  • RNG105/caprin1, an RNA granule protein for dendritic mRNA localization, is essential for long-term memory formation. 査読 国際誌

    Kei Nakayama, Rie Ohashi, Yo Shinoda, Maya Yamazaki, Manabu Abe, Akihiro Fujikawa, Shuji Shigenobu, Akira Futatsugi, Masaharu Noda, Katsuhiko Mikoshiba, Teiichi Furuichi, Kenji Sakimura, Nobuyuki Shiina

    eLife   6   2017年11月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Local regulation of synaptic efficacy is thought to be important for proper networking of neurons and memory formation. Dysregulation of global translation influences long-term memory in mice, but the relevance of the regulation specific for local translation by RNA granules remains elusive. Here, we demonstrate roles of RNG105/caprin1 in long-term memory formation. RNG105 deletion in mice impaired synaptic strength and structural plasticity in hippocampal neurons. Furthermore, RNG105-deficient mice displayed unprecedentedly severe defects in long-term memory formation in spatial and contextual learning tasks. Genome-wide profiling of mRNA distribution in the hippocampus revealed an underlying mechanism: RNG105 deficiency impaired the asymmetric somato-dendritic localization of mRNAs. Particularly, RNG105 deficiency reduced the dendritic localization of mRNAs encoding regulators of AMPAR surface expression, which was consistent with attenuated homeostatic AMPAR scaling in dendrites and reduced synaptic strength. Thus, RNG105 has an essential role, as a key regulator of dendritic mRNA localization, in long-term memory formation.

    DOI: 10.7554/eLife.29677

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  • Orexin modulates behavioral fear expression through the locus coeruleus. 査読 国際誌

    Shingo Soya, Tohru M Takahashi, Thomas J McHugh, Takashi Maejima, Stefan Herlitze, Manabu Abe, Kenji Sakimura, Takeshi Sakurai

    Nature communications   8 ( 1 )   1606 - 1606   2017年11月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:NATURE PUBLISHING GROUP  

    Emotionally salient information activates orexin neurons in the lateral hypothalamus, leading to increase in sympathetic outflow and vigilance level. How this circuit alters animals' behavior remains unknown. Here we report that noradrenergic neurons in the locus coeruleus (NALC neurons) projecting to the lateral amygdala (LA) receive synaptic input from orexin neurons. Pharmacogenetic/optogenetic silencing of this circuit as well as acute blockade of the orexin receptor-1 (OX1R) decreases conditioned fear responses. In contrast, optogenetic stimulation of this circuit potentiates freezing behavior against a similar but distinct context or cue. Increase of orexinergic tone by fasting also potentiates freezing behavior and LA activity, which are blocked by pharmacological blockade of OX1R in the LC. These findings demonstrate the circuit involving orexin, NALC and LA neurons mediates fear-related behavior and suggests inappropriate excitation of this pathway may cause fear generalization sometimes seen in psychiatric disorders, such as PTSD.

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  • The number and distribution of AMPA receptor channels containing fast kinetic GluA3 and GluA4 subunits at auditory nerve synapses depend on the target cells. 査読 国際誌

    María E Rubio, Ko Matsui, Yugo Fukazawa, Naomi Kamasawa, Harumi Harada, Makoto Itakura, Elek Molnár, Manabu Abe, Kenji Sakimura, Ryuichi Shigemoto

    Brain structure & function   222 ( 8 )   3375 - 3393   2017年11月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:SPRINGER HEIDELBERG  

    The neurotransmitter receptor subtype, number, density, and distribution relative to the location of transmitter release sites are key determinants of signal transmission. AMPA-type ionotropic glutamate receptors (AMPARs) containing GluA3 and GluA4 subunits are prominently expressed in subsets of neurons capable of firing action potentials at high frequencies, such as auditory relay neurons. The auditory nerve (AN) forms glutamatergic synapses on two types of relay neurons, bushy cells (BCs) and fusiform cells (FCs) of the cochlear nucleus. AN-BC and AN-FC synapses have distinct kinetics; thus, we investigated whether the number, density, and localization of GluA3 and GluA4 subunits in these synapses are differentially organized using quantitative freeze-fracture replica immunogold labeling. We identify a positive correlation between the number of AMPARs and the size of AN-BC and AN-FC synapses. Both types of AN synapses have similar numbers of AMPARs; however, the AN-BC have a higher density of AMPARs than AN-FC synapses, because the AN-BC synapses are smaller. A higher number and density of GluA3 subunits are observed at AN-BC synapses, whereas a higher number and density of GluA4 subunits are observed at AN-FC synapses. The intrasynaptic distribution of immunogold labeling revealed that AMPAR subunits, particularly GluA3, are concentrated at the center of the AN-BC synapses. The central distribution of AMPARs is absent in GluA3-knockout mice, and gold particles are evenly distributed along the postsynaptic density. GluA4 gold labeling was homogenously distributed along both synapse types. Thus, GluA3 and GluA4 subunits are distributed at AN synapses in a target-cell-dependent manner.

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  • Netrin-1 Derived from the Ventricular Zone, but not the Floor Plate, Directs Hindbrain Commissural Axons to the Ventral Midline. 査読 国際誌

    Kenta Yamauchi, Maya Yamazaki, Manabu Abe, Kenji Sakimura, Heiko Lickert, Takahiko Kawasaki, Fujio Murakami, Tatsumi Hirata

    Scientific reports   7 ( 1 )   11992 - 11992   2017年9月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:NATURE PUBLISHING GROUP  

    Netrin-1 (Ntn1) emanating from the ventral midline has been thought to act as a long-range diffusible chemoattractant for commissural axons (CAs). However, CAs still grow towards the midline in the absence of the floor plate (FP), a glial structure occupying the midline. Here, using genetically loss-of-function approaches in mice, we show that Ntn1 derived from the ventricular zone (VZ), but not the FP, is crucial for CA guidance in the mouse hindbrain. During the period of CA growth, Ntn1 is expressed in the ventral two-thirds of the VZ, in addition to the FP. Remarkably, deletion of Ntn1 from the VZ and even from the dorsal VZ highly disrupts CA guidance to the midline, whereas the deletion from the FP has little impact on it. We also show that the severities of CA guidance defects found in the Ntn1 conditional mutants were irrelevant to their FP long-range chemoattractive activities. Our results are incompatible with the prevailing view that Ntn1 is an FP-derived long-range diffusible chemoattractant for CAs, but suggest a novel mechanism that VZ-derived Ntn1 directs CAs to the ventral midline by its local actions.

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  • An RNA-binding protein, Qki5, regulates embryonic neural stem cells through pre-mRNA processing in cell adhesion signaling. 査読 国際誌

    Yoshika Hayakawa-Yano, Satoshi Suyama, Masahiro Nogami, Masato Yugami, Ikuko Koya, Takako Furukawa, Li Zhou, Manabu Abe, Kenji Sakimura, Hirohide Takebayashi, Atsushi Nakanishi, Hideyuki Okano, Masato Yano

    Genes & development   31 ( 18 )   1910 - 1925   2017年9月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:COLD SPRING HARBOR LAB PRESS, PUBLICATIONS DEPT  

    Cell type-specific transcriptomes are enabled by the action of multiple regulators, which are frequently expressed within restricted tissue regions. In the present study, we identify one such regulator, Quaking 5 (Qki5), as an RNA-binding protein (RNABP) that is expressed in early embryonic neural stem cells and subsequently down-regulated during neurogenesis. mRNA sequencing analysis in neural stem cell culture indicates that Qki proteins play supporting roles in the neural stem cell transcriptome and various forms of mRNA processing that may result from regionally restricted expression and subcellular localization. Also, our in utero electroporation gain-of-function study suggests that the nuclear-type Qki isoform Qki5 supports the neural stem cell state. We next performed in vivo transcriptome-wide protein-RNA interaction mapping to search for direct targets of Qki5 and elucidate how Qki5 regulates neural stem cell function. Combined with our transcriptome analysis, this mapping analysis yielded a bona fide map of Qki5-RNA interaction at single-nucleotide resolution, the identification of 892 Qki5 direct target genes, and an accurate Qki5-dependent alternative splicing rule in the developing brain. Last, our target gene list provides the first compelling evidence that Qki5 is associated with specific biological events; namely, cell-cell adhesion. This prediction was confirmed by histological analysis of mice in which Qki proteins were genetically ablated, which revealed disruption of the apical surface of the lateral wall in the developing brain. These data collectively indicate that Qki5 regulates communication between neural stem cells by mediating numerous RNA processing events and suggest new links between splicing regulation and neural stem cell states.

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  • Unconventional role of voltage-gated proton channels (VSOP/Hv1) in regulation of microglial ROS production. 査読 国際誌

    Takafumi Kawai, Yoshifumi Okochi, Tomohiko Ozaki, Yoshio Imura, Schuichi Koizumi, Maya Yamazaki, Manabu Abe, Kenji Sakimura, Toshihide Yamashita, Yasushi Okamura

    Journal of neurochemistry   142 ( 5 )   686 - 699   2017年9月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:WILEY  

    It has been established that voltage-gated proton channels (VSOP/Hv1), encoded by Hvcn1, support reactive oxygen species (ROS) production in phagocytic activities of neutrophils (El Chemaly et al. ) and antibody production in B lymphocytes (Capasso et al. ). VSOP/Hv1 is a potential therapeutic target for brain ischemia, since Hvcn1 deficiency reduces microglial ROS production and protects brain from neuronal damage (Wu et al. ). In the present study, we report that VSOP/Hv1 has paradoxical suppressive role in ROS production in microglia. Extracellular ROS production was lower in neutrophils of Hvcn1-/- mice than WT mice as reported. In contrast, it was drastically enhanced in isolated Hvcn1-/- microglia as compared with cells from WT mice. Actin dynamics was altered in Hvcn1-/- microglia and intracellular distribution of cytosolic NADPH oxidase subunit, p67, was changed. When expression levels of oxidative stress responsive antioxidant genes were compared between WT and Hvcn1-/- in cerebral cortex at different ages of animals, they were slightly decreased in Hvcn1-/- mice at younger stage (1 day, 5 days, 3 weeks old), but drastically increased at aged stage (6 months old), suggesting that the regulation of microglial ROS production by VSOP/Hv1 is age-dependent. We also performed brain ischemic stroke experiments and found that the neuroprotective effect of VSOP/Hv1deficiency on infarct volume depended on the age of animals. Taken together, regulation of ROS production by VSOP/Hv1 is more complex than previously thought and significance of VSOP/Hv1 in microglial ROS production depends on age.

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  • Impaired lymphocyte trafficking in mice deficient in the kinase activity of PKN1. 査読 国際誌

    Rana Mashud, Akira Nomachi, Akihide Hayakawa, Koji Kubouchi, Sally Danno, Takako Hirata, Kazuhiko Matsuo, Takashi Nakayama, Ryosuke Satoh, Reiko Sugiura, Manabu Abe, Kenji Sakimura, Shigeharu Wakana, Hiroyuki Ohsaki, Shingo Kamoshida, Hideyuki Mukai

    Scientific reports   7 ( 1 )   7663 - 7663   2017年8月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:NATURE PUBLISHING GROUP  

    Knock-in mice lacking PKN1 kinase activity were generated by introducing a T778A point mutation in the catalytic domain. PKN1[T778A] mutant mice developed to adulthood without apparent external abnormalities, but exhibited lower T and B lymphocyte counts in the peripheral blood than those of wild-type (WT) mice. T and B cell development proceeded in an apparently normal fashion in bone marrow and thymus of PKN1[T778A] mice, however, the number of T and B cell counts were significantly higher in the lymph nodes and spleen of mutant mice in those of WT mice. After transfusion into WT recipients, EGFP-labelled PKN1[T778A] donor lymphocytes were significantly less abundant in the peripheral circulation and more abundant in the spleen and lymph nodes of recipient mice compared with EGFP-labelled WT donor lymphocytes, likely reflecting lymphocyte sequestration in the spleen and lymph nodes in a cell-autonomous fashion. PKN1[T778A] lymphocytes showed significantly lower chemotaxis towards chemokines and sphingosine 1-phosphate (S1P) than WT cells in vitro. The biggest migration defect was observed in response to S1P, which is essential for lymphocyte egress from secondary lymphoid organs. These results reveal a novel role of PKN1 in lymphocyte migration and localization.

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  • The balance between cathepsin C and cystatin F controls remyelination in the brain of Plp1-overexpressing mouse, a chronic demyelinating disease model. 査読 国際誌

    Takahiro Shimizu, Wilaiwan Wisessmith, Jiayi Li, Manabu Abe, Kenji Sakimura, Banthit Chetsawang, Yoshinori Sahara, Koujiro Tohyama, Kenji F Tanaka, Kazuhiro Ikenaka

    Glia   65 ( 6 )   917 - 930   2017年6月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:WILEY  

    In demyelinating diseases such as multiple sclerosis (MS), an imbalance between the demyelination and remyelination rates underlies the degenerative processes. Microglial activation is observed in demyelinating lesions; however, the molecular mechanism responsible for the homeostatic/environmental change remains elusive. We previously found that cystatin F (CysF), a cysteine protease inhibitor, is selectively expressed in microglia only in actively demyelinating/remyelinating lesions but ceases expression in chronic lesions, suggesting its role in remyelination. Here, we report the effects of manipulating the expression of CysF and cathepsin C (CatC), a key target of CysF, in a murine model of transgenic demyelinating disease, Plp4e/- . During the active remyelinating phase, both CysF knockdown (CysFKD) and microglial-selective CatC overexpression (CatCOE) showed a worsening of the demyelination in Plp4e/- transgenic mice. Conversely, during the chronic demyelinating phase, CatC knockdown (CatCKD) ameliorated the demyelination. Our results suggest that the balance between CatC and CysF expression controls the demyelination and remyelination process.

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  • Distribution of corticotropin-releasing factor neurons in the mouse brain: a study using corticotropin-releasing factor-modified yellow fluorescent protein knock-in mouse. 査読 国際誌

    Junko Kono, Kohtarou Konno, Ashraf Hossain Talukder, Toshimitsu Fuse, Manabu Abe, Katsuya Uchida, Shuhei Horio, Kenji Sakimura, Masahiko Watanabe, Keiichi Itoi

    Brain structure & function   222 ( 4 )   1705 - 1732   2017年5月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:SPRINGER HEIDELBERG  

    We examined the morphological features of corticotropin-releasing factor (CRF) neurons in a mouse line in which modified yellow fluorescent protein (Venus) was expressed under the CRF promoter. We previously generated the CRF-Venus knock-in mouse, in which Venus is inserted into the CRF gene locus by homologous recombination. In the present study, the neomycin phosphotransferase gene (Neo), driven by the pgk-1 promoter, was deleted from the CRF-Venus mouse genome, and a CRF-Venus∆Neo mouse was generated. Venus expression is much more prominent in the CRF-Venus∆Neo mouse when compared to the CRF-Venus mouse. In addition, most Venus-expressing neurons co-express CRF mRNA. Venus-expressing neurons constitute a discrete population of neuroendocrine neurons in the paraventricular nucleus of the hypothalamus (PVH) that project to the median eminence. Venus-expressing neurons were also found in brain regions outside the neuroendocrine PVH, including the olfactory bulb, the piriform cortex (Pir), the extended amygdala, the hippocampus, the neocortices, Barrington's nucleus, the midbrain/pontine dorsal tegmentum, the periaqueductal gray, and the inferior olivary nucleus (IO). Venus-expressing perikarya co-expressing CRF mRNA could be observed clearly even in regions where CRF-immunoreactive perikarya could hardly be identified. We demonstrated that the CRF neurons contain glutamate in the Pir and IO, while they contain gamma-aminobutyric acid in the neocortex, the bed nucleus of the stria terminalis, the hippocampus, and the amygdala. A population of CRF neurons was demonstrated to be cholinergic in the midbrain tegmentum. The CRF-Venus∆Neo mouse may be useful for studying the structural and functional properties of CRF neurons in the mouse brain.

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  • YAP functions as a mechanotransducer in oligodendrocyte morphogenesis and maturation. 査読 国際誌

    Takeshi Shimizu, Yasuyuki Osanai, Kenji F Tanaka, Manabu Abe, Rie Natsume, Kenji Sakimura, Kazuhiro Ikenaka

    Glia   65 ( 2 )   360 - 374   2017年2月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:WILEY-BLACKWELL  

    Oligodendrocytes (OLs) are myelinating cells of the central nervous system. Recent studies have shown that mechanical factors influence various cell properties. Mechanical stimuli can be transduced into intracellular biochemical signals through mechanosensors and intracellular mechanotransducers, such as YAP. However, the molecular mechanisms underlying mechanical regulation of OLs by YAP remain unknown. We found that OL morphology and interactions between OLs and neuronal axons were affected by knocking down YAP. Mechanical stretching of OL precursor cells induced nuclear YAP accumulation and assembly of focal adhesion, key platforms for mechanotransduction. Shear stress decreased the number of OL processes, whereas a dominant-negative form of YAP suppressed these effects. To investigate the roles of YAP in postnatal OLs in vivo, we constructed a novel YAP knock-in mouse and found that in vivo overexpression of YAP widely affected OL maturation. These results indicate that YAP regulates OL morphology and maturation in response to mechanical factors. GLIA 2017;65:360-374.

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  • PKN2 is essential for mouse embryonic development and proliferation of mouse fibroblasts. 査読 国際誌

    Sally Danno, Koji Kubouchi, Mona Mehruba, Manabu Abe, Rie Natsume, Kenji Sakimura, Satoshi Eguchi, Masahiro Oka, Masanori Hirashima, Hiroki Yasuda, Hideyuki Mukai

    Genes to cells : devoted to molecular & cellular mechanisms   22 ( 2 )   220 - 236   2017年2月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:WILEY  

    PKN2, a member of the protein kinase N (PKN) family, has been suggested by in vitro culture cell experiments to bind to Rho/Rac GTPases and contributes to cell-cell contact and cell migration. To unravel the in vivo physiological function of PKN2, we targeted the PKN2 gene. Constitutive disruption of the mouse PKN2 gene resulted in growth retardation and lethality before embryonic day (E) 10.5. PKN2-/- embryo did not undergo axial turning and showed insufficient closure of the neural tube. Mouse embryonic fibroblasts (MEFs) derived from PKN2-/- embryos at E9.5 failed to grow. Cre-mediated ablation of PKN2 in PKN2flox/flox MEFs obtained from E14.5 embryos showed impaired cell proliferation, and cell cycle analysis of these MEFs showed a decrease in S-phase population. Our results show that PKN2 is essential for mouse embryonic development and cell-autonomous proliferation of primary MEFs in culture. Comparison of the PKN2-/- phenotype with the phenotypes of PKN1 and PKN3 knockout strains suggests that PKN2 has distinct nonredundant functions in vivo, despite the structural similarity and evolutionary relationship among the three isoforms.

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  • Impaired auditory processing and altered structure of the endbulb of Held synapse in mice lacking the GluA3 subunit of AMPA receptors. 査読 国際誌

    Sofía García-Hernández, Manabu Abe, Kenji Sakimura, María E Rubio

    Hearing research   344   284 - 294   2017年2月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ELSEVIER SCIENCE BV  

    AMPA glutamate receptor complexes with fast kinetics conferred by subunits like GluA3 and GluA4 are essential for temporal precision of synaptic transmission. The specific role of GluA3 in auditory processing and experience related changes in the auditory brainstem remain unknown. We investigated the role of the GluA3 in auditory processing by using wild type (WT) and GluA3 knockout (GluA3-KO) mice. We recorded auditory brainstem responses (ABR) to assess auditory function and used electron microscopy to evaluate the ultrastructure of the auditory nerve synapse on bushy cells (AN-BC synapse). Since labeling for GluA3 subunit increases on auditory nerve synapses within the cochlear nucleus in response to transient sound reduction, we investigated the role of GluA3 in experience-dependent changes in auditory processing. We induced transient sound reduction by plugging one ear and evaluated ABR threshold and peak amplitude recovery for up to 60 days after ear plug removal in WT and GluA3-KO mice. We found that the deletion of GluA3 leads to impaired auditory signaling that is reflected in decreased ABR peak amplitudes, an increased latency of peak 2, early onset hearing loss and reduced numbers and sizes of postsynaptic densities (PSDs) of AN-BC synapses. Additionally, the lack of GluA3 hampers ABR threshold recovery after transient ear plugging. We conclude that GluA3 is required for normal auditory signaling, normal ultrastructure of AN-BC synapses in the cochlear nucleus and normal experience-dependent changes in auditory processing after transient sound reduction.

    DOI: 10.1016/j.heares.2016.12.006

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  • Forward-genetics analysis of sleep in randomly mutagenized mice. 査読 国際誌

    Hiromasa Funato, Chika Miyoshi, Tomoyuki Fujiyama, Takeshi Kanda, Makito Sato, Zhiqiang Wang, Jing Ma, Shin Nakane, Jun Tomita, Aya Ikkyu, Miyo Kakizaki, Noriko Hotta-Hirashima, Satomi Kanno, Haruna Komiya, Fuyuki Asano, Takato Honda, Staci J Kim, Kanako Harano, Hiroki Muramoto, Toshiya Yonezawa, Seiya Mizuno, Shinichi Miyazaki, Linzi Connor, Vivek Kumar, Ikuo Miura, Tomohiro Suzuki, Atsushi Watanabe, Manabu Abe, Fumihiro Sugiyama, Satoru Takahashi, Kenji Sakimura, Yu Hayashi, Qinghua Liu, Kazuhiko Kume, Shigeharu Wakana, Joseph S Takahashi, Masashi Yanagisawa

    Nature   539 ( 7629 )   378 - 383   2016年11月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:NATURE PUBLISHING GROUP  

    Sleep is conserved from invertebrates to vertebrates, and is tightly regulated in a homeostatic manner. The molecular and cellular mechanisms that determine the amount of rapid eye movement sleep (REMS) and non-REMS (NREMS) remain unknown. Here we identify two dominant mutations that affect sleep and wakefulness by using an electroencephalogram/electromyogram-based screen of randomly mutagenized mice. A splicing mutation in the Sik3 protein kinase gene causes a profound decrease in total wake time, owing to an increase in inherent sleep need. Sleep deprivation affects phosphorylation of regulatory sites on the kinase, suggesting a role for SIK3 in the homeostatic regulation of sleep amount. Sik3 orthologues also regulate sleep in fruitflies and roundworms. A missense, gain-of-function mutation in the sodium leak channel NALCN reduces the total amount and episode duration of REMS, apparently by increasing the excitability of REMS-inhibiting neurons. Our results substantiate the use of a forward-genetics approach for studying sleep behaviours in mice, and demonstrate the role of SIK3 and NALCN in regulating the amount of NREMS and REMS, respectively.

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  • Roles of Cbln1 in Non-Motor Functions of Mice. 査読 国際誌

    Shintaro Otsuka, Kohtarou Konno, Manabu Abe, Junko Motohashi, Kazuhisa Kohda, Kenji Sakimura, Masahiko Watanabe, Michisuke Yuzaki

    The Journal of neuroscience : the official journal of the Society for Neuroscience   36 ( 46 )   11801 - 11816   2016年11月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:SOC NEUROSCIENCE  

    The cerebellum is thought to be involved in cognitive functions in addition to its well established role in motor coordination and motor learning in humans. Cerebellin 1 (Cbln1) is predominantly expressed in cerebellar granule cells and plays a crucial role in the formation and function of parallel fiber-Purkinje cell synapses. Although genes encoding Cbln1 and its postsynaptic receptor, the delta2 glutamate receptor (GluD2), are suggested to be associated with autistic-like traits and many psychiatric disorders, whether such cognitive impairments are caused by cerebellar dysfunction remains unclear. In the present study, we investigated whether and how Cbln1 signaling is involved in non-motor functions in adult mice. We show that acquisition and retention/retrieval of cued and contextual fear memory were impaired in Cbln1-null mice. In situ hybridization and immunohistochemical analyses revealed that Cbln1 is expressed in various extracerebellar regions, including the retrosplenial granular cortex and the hippocampus. In the hippocampus, Cbln1 immunoreactivity was present at the molecular layer of the dentate gyrus and the stratum lacunosum-moleculare without overt mRNA expression, suggesting that Cbln1 is provided by perforant path fibers. Retention/retrieval, but not acquisition, of cued and contextual fear memory was impaired in forebrain-predominant Cbln1-null mice. Spatial learning in the radial arm water maze was also abrogated. In contrast, acquisition of fear memory was affected in cerebellum-predominant Cbln1-null mice. These results indicate that Cbln1 in the forebrain and cerebellum mediates specific aspects of fear conditioning and spatial memory differentially and that Cbln1 signaling likely regulates motor and non-motor functions in multiple brain regions. SIGNIFICANCE STATEMENT: Despites its well known role in motor coordination and motor learning, whether and how the cerebellum is involved in cognitive functions remains less clear. Cerebellin 1 (Cbln1) is highly expressed in the cerebellum and serves as an essential synaptic organizer. Although genes encoding Cbln1 and its receptor are associated with many psychiatric disorders, it remains unknown whether such cognitive impairments are caused by cerebellar dysfunction. Here, we show that Cbln1 is also expressed in the forebrain, including the hippocampus and retrosplenial granular cortex. Using forebrain- and cerebellum-predominant conditional Cbln1-null mice, we show that Cbln1 in the forebrain and cerebellum mediates specific aspects of fear conditioning and spatial memory differentially, indicating that Cbln1 signaling regulates both motor and non-motor functions in multiple brain regions.

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  • SAD-B Phosphorylation of CAST Controls Active Zone Vesicle Recycling for Synaptic Depression. 査読 国際誌

    Sumiko Mochida, Yamato Hida, Shota Tanifuji, Akari Hagiwara, Shun Hamada, Manabu Abe, Huan Ma, Misato Yasumura, Isao Kitajima, Kenji Sakimura, Toshihisa Ohtsuka

    Cell reports   16 ( 11 )   2901 - 2913   2016年9月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:CELL PRESS  

    Short-term synaptic depression (STD) is a common form of activity-dependent plasticity observed widely in the nervous system. Few molecular pathways that control STD have been described, but the active zone (AZ) release apparatus provides a possible link between neuronal activity and plasticity. Here, we show that an AZ cytomatrix protein CAST and an AZ-associated protein kinase SAD-B coordinately regulate STD by controlling reloading of the AZ with release-ready synaptic vesicles. SAD-B phosphorylates the N-terminal serine (S45) of CAST, and S45 phosphorylation increases with higher firing rate. A phosphomimetic CAST (S45D) mimics CAST deletion, which enhances STD by delaying reloading of the readily releasable pool (RRP), resulting in a pool size decrease. A phosphonegative CAST (S45A) inhibits STD and accelerates RRP reloading. Our results suggest that the CAST/SAD-B reaction serves as a brake on synaptic transmission by temporal calibration of activity and synaptic depression via RRP size regulation.

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  • Involvement of Brain-Enriched guanylate Kinase-Associated protein (BEGAIN) in chronic pain after peripheral nerve injury 査読

    Tayo Katano, Masafumi Fukuda, Hidemasa Furue, Maya Yamazaki, Manabu Abe, Masahiko Watanabe, Kazuhiko Nishida, Ikuko Yao, Akihiro Yamada, Yutaka Hata, Nobuaki Okumura, Takanobu Nakazawa, Tadashi Yamamoto, Kenji Sakimura, Toshifumi Takao, Seiji Ito

    eNeuro   3 ( 5 )   2016年9月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:Society for Neuroscience  

    Maintenance of neuropathic pain caused by peripheral nerve injury crucially depends on the phosphorylation of GluN2B, a subunit of the N-methyl-d-aspartate (NMDA) receptor, at Tyr1472 (Y1472) and subsequent formation of a postsynaptic density (PSD) complex of superficial spinal dorsal horn neurons. Here we took advantage of comparative proteomic analysis based on isobaric stable isotope tags (iTRAQ) between wild-type and knock-in mice with a mutation of Y1472 to Phe of GluN2B (Y1472F-KI) to search for PSD proteins in the spinal dorsal horn that mediate the signaling downstream of phosphorylated Y1472 GluN2B. Among several candidate proteins, we focused on brain-enriched guanylate kinase-associated protein (BEGAIN), which was specifically up-regulated in wild-type mice after spared nerve injury (SNI). Immunohistochemical analysis using the generated antibody demonstrated that BEGAIN was highly localized at the synapse of inner lamina II in the spinal dorsal horn and that its expression was up-regulated after SNI in wild-type, but not in Y1472F-KI, mice. In addition, alteration of the kinetics of evoked excitatory postsynaptic currents for NMDA but not those for _-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptors in spinal lamina II was demonstrated by BEGAIN deletion. We demonstrated that mechanical allodynia, a condition of abnormal pain induced by innocuous stimuli, in the SNI model was significantly attenuated in BEGAIN-deficient mice. However, there was no significant difference between naive wild-type and BEGAIN-knockout mice in terms of physiological threshold for mechanical stimuli. These results suggest that BEGAIN was involved in pathological pain transmission through NMDA receptor activation by the phosphorylation of GluN2B at Y1472 in spinal inner lamina II.

    DOI: 10.1523/ENEURO.0110-16.2016

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  • Ttll9-/- mice sperm flagella show shortening of doublet 7, reduction of doublet 5 polyglutamylation and a stall in beating. 査読 国際誌

    Alu Konno, Koji Ikegami, Yoshiyuki Konishi, Hyun-Jeong Yang, Manabu Abe, Maya Yamazaki, Kenji Sakimura, Ikuko Yao, Kogiku Shiba, Kazuo Inaba, Mitsutoshi Setou

    Journal of cell science   129 ( 14 )   2757 - 66   2016年7月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:COMPANY OF BIOLOGISTS LTD  

    Nine outer doublet microtubules in axonemes of flagella and cilia are heterogeneous in structure and biochemical properties. In mammalian sperm flagella, one of the factors to generate the heterogeneity is tubulin polyglutamylation, although the importance of the heterogeneous modification is unclear. Here, we show that a tubulin polyglutamylase Ttll9 deficiency (Ttll9(-/-)) causes a unique set of phenotypes related to doublet heterogeneity. Ttll9(-/-) sperm axonemes had frequent loss of a doublet and reduced polyglutamylation. Intriguingly, the doublet loss selectively occurred at the distal region of doublet 7, and reduced polyglutamylation was observed preferentially on doublet 5. Ttll9(-/-) spermatozoa showed aberrant flagellar beating, characterized by frequent stalls after anti-hook bending. This abnormal motility could be attributed to the reduction of polyglutamylation on doublet 5, which probably occurred at a position involved in the switching of bending. These results indicate that mammalian Ttll9 plays essential roles in maintaining the normal structure and beating pattern of sperm flagella by establishing normal heterogeneous polyglutamylation patterns.

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  • Transsynaptic Modulation of Kainate Receptor Functions by C1q-like Proteins. 査読 国際誌

    Keiko Matsuda, Timotheus Budisantoso, Nikolaos Mitakidis, Yuki Sugaya, Eriko Miura, Wataru Kakegawa, Miwako Yamasaki, Kohtarou Konno, Motokazu Uchigashima, Manabu Abe, Izumi Watanabe, Masanobu Kano, Masahiko Watanabe, Kenji Sakimura, A Radu Aricescu, Michisuke Yuzaki

    Neuron   90 ( 4 )   752 - 67   2016年5月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:CELL PRESS  

    Postsynaptic kainate-type glutamate receptors (KARs) regulate synaptic network activity through their slow channel kinetics, most prominently at mossy fiber (MF)-CA3 synapses in the hippocampus. Nevertheless, how KARs cluster and function at these synapses has been unclear. Here, we show that C1q-like proteins C1ql2 and C1ql3, produced by MFs, serve as extracellular organizers to recruit functional postsynaptic KAR complexes to the CA3 pyramidal neurons. C1ql2 and C1ql3 specifically bound the amino-terminal domains of postsynaptic GluK2 and GluK4 KAR subunits and the presynaptic neurexin 3 containing a specific sequence in vitro. In C1ql2/3 double-null mice, CA3 synaptic responses lost the slow, KAR-mediated components. Furthermore, despite induction of MF sprouting in a temporal lobe epilepsy model, KARs were not recruited to postsynaptic sites in C1ql2/3 double-null mice, leading to reduced recurrent circuit activities. C1q family proteins, broadly expressed, are likely to modulate KAR function throughout the brain and represent promising antiepileptic targets.

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  • TARP γ-2 and γ-8 Differentially Control AMPAR Density Across Schaffer Collateral/Commissural Synapses in the Hippocampal CA1 Area. 査読 国際誌

    Yamasaki M, Fukaya M, Yamazaki M, Azechi H, Natsume R, Abe M, Sakimura K, Watanabe M

    The Journal of neuroscience : the official journal of the Society for Neuroscience   36 ( 15 )   4296 - 312   2016年4月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    DOI: 10.1523/JNEUROSCI.4178-15.2016

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  • Determination of kainate receptor subunit ratios in mouse brain using novel chimeric protein standards. 査読 国際誌

    Izumi Watanabe-Iida, Kohtarou Konno, Kaori Akashi, Manabu Abe, Rie Natsume, Masahiko Watanabe, Kenji Sakimura

    Journal of neurochemistry   136 ( 2 )   295 - 305   2016年1月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:WILEY-BLACKWELL  

    Kainate-type glutamate receptors (KARs) are tetrameric channels assembled from GluK1-5. GluK1-3 are low-affinity subunits that form homomeric and heteromeric KARs, while GluK4 and GluK5 are high-affinity subunits that require co-assembly with GluK1-3 for functional expression. Although the subunit composition is thought to be highly heterogeneous in the brain, the distribution of KAR subunits at the protein level and their relative abundance in given regions of the brain remain largely unknown. In the present study, we titrated C-terminal antibodies to each KAR subunit using chimeric GluA2-GluK fusion proteins, and measured their relative abundance in the P2 and post-synaptic density (PSD) fractions of the adult mouse hippocampus and cerebellum. Analytical western blots showed that GluK2 and GluK3 were the major KAR subunits, with additional expression of GluK5 in the hippocampus and cerebellum. In both regions, GluK4 was very low and GluK1 was below the detection threshold. The relative amount of low-affinity subunits (GluK2 plus GluK3) was several times higher than that of high-affinity subunits (GluK4 plus GluK5) in both regions. Of note, the highest ratio of high-affinity subunits to low-affinity subunits was found in the hippocampal PSD fraction (0.32), suggesting that heteromeric receptors consisting of high- and low-affinity subunits highly accumulate at hippocampal synapses. In comparison, this ratio was decreased to 0.15 in the cerebellar PSD fraction, suggesting that KARs consisting of low-affinity subunits are more prevalent in the cerebellum. Therefore, low-affinity KAR subunits are predominant in the brain, with distinct subunit combinations between the hippocampus and cerebellum. Kainate receptors, an unconventional member of the iGluR receptor family, have a tetrameric structure assembled from low-affinity (GluK1-3) and high-affinity (GluK4 and GluK5) subunits. We used a simple but novel procedure to measure the relative abundance of both low- and high-affinity subunits. This method revealed that the relative amount of GluK2 plus GluK3 subunits was several times higher than that of GluK4 plus GluK5 subunits, in both the hippocampus and cerebellum.

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  • A CDC42EP4/septin-based perisynaptic glial scaffold facilitates glutamate clearance. 査読 国際誌

    Natsumi Ageta-Ishihara, Maya Yamazaki, Kohtarou Konno, Hisako Nakayama, Manabu Abe, Kenji Hashimoto, Tomoki Nishioka, Kozo Kaibuchi, Satoko Hattori, Tsuyoshi Miyakawa, Kohichi Tanaka, Fathul Huda, Hirokazu Hirai, Kouichi Hashimoto, Masahiko Watanabe, Kenji Sakimura, Makoto Kinoshita

    Nature communications   6   10090 - 10090   2015年12月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    The small GTPase-effector proteins CDC42EP1-5/BORG1-5 interact reciprocally with CDC42 or the septin cytoskeleton. Here we show that, in the cerebellum, CDC42EP4 is exclusively expressed in Bergmann glia and localizes beneath specific membrane domains enwrapping dendritic spines of Purkinje cells. CDC42EP4 forms complexes with septin hetero-oligomers, which interact with a subset of glutamate transporter GLAST/EAAT1. In Cdc42ep4(-/-) mice, GLAST is dissociated from septins and is delocalized away from the parallel fibre-Purkinje cell synapses. The excitatory postsynaptic current exhibits a protracted decay time constant, reduced sensitivity to a competitive inhibitor of the AMPA-type glutamate receptors (γDGG) and excessive baseline inward current in response to a subthreshold dose of a nonselective inhibitor of the glutamate transporters/EAAT1-5 (DL-TBOA). Insufficient glutamate-buffering/clearance capacity in these mice manifests as motor coordination/learning defects, which are aggravated with subthreshold DL-TBOA. We propose that the CDC42EP4/septin-based glial scaffold facilitates perisynaptic localization of GLAST and optimizes the efficiency of glutamate-buffering and clearance.

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  • Corrigendum to "Transient expression of neuropeptide W in postnatal mouse hypothalamus - A putative regulator of energy homeostasis" [Neuroscience, 301 (2015) 323-337] 査読

    T. Motoike, A. G. Skach, J. K. Godwin, C. M. Sinton, M. Yamazaki, M. Abe, R. Natsume, K. Sakimura, M. Yanagisawa

    Neuroscience   303   630   2015年9月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:Elsevier Ltd  

    DOI: 10.1016/j.neuroscience.2015.06.064

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  • STAT5 Orchestrates Local Epigenetic Changes for Chromatin Accessibility and Rearrangements by Direct Binding to the TCRγ Locus. 査読 国際誌

    Wagatsuma K, Tani-ichi S, Liang B, Shitara S, Ishihara K, Abe M, Miyachi H, Kitano S, Hara T, Nanno M, Ishikawa H, Sakimura K, Nakao M, Kimura H, Ikuta K

    Journal of immunology (Baltimore, Md. : 1950)   195 ( 4 )   1804 - 14   2015年8月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    DOI: 10.4049/jimmunol.1302456

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  • Horizontal Basal Cell-Specific Deletion of Pax6 Impedes Recovery of the Olfactory Neuroepithelium Following Severe Injury. 査読 国際誌

    Jun Suzuki, Katsuyasu Sakurai, Maya Yamazaki, Manabu Abe, Hitoshi Inada, Kenji Sakimura, Yukio Katori, Noriko Osumi

    Stem cells and development   24 ( 16 )   1923 - 33   2015年8月

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    記述言語:英語  

    In the mammalian olfactory epithelium (OE), olfactory receptor neurons (ORNs) are continuously regenerated throughout the animal's lifetime. Horizontal basal cells (HBCs) in the OE express the epithelial marker keratin 5 (K5) and the stem cell marker Pax6 and are considered relatively quiescent tissue stem cells in the OE. Pax6 is a key regulator of several developmental processes in the central nervous system and in sensory organs. Although Pax6 is expressed in the OE, its precise role remains unknown, particularly with respect to stem cell-like HBCs. To investigate the function of Pax6 in the developmental and regenerative processes in the OE, we generated conditional Pax6-knockout mice carrying a loxP-floxed Pax6 gene. Homozygous Pax6-floxed mice were crossed with K5-Cre transgenic mice to generate HBC-specific Pax6-knockout (Pax6-cKO) mice. We confirmed that the deletion of Pax6 expression in HBCs was sufficiently achieved in zone 1 of the OE in Pax6-cKO mice 3 days after methimazole-induced severe damage. In this condition, regeneration of the OE was dramatically impaired; both OE thickness and the number of ORNs were significantly decreased in the regenerated OE of Pax6-cKO mice. These results suggest that Pax6 expression is essential for HBCs to differentiate into neuronal cells during the regeneration process following severe injury.

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  • TRANSIENT EXPRESSION OF NEUROPEPTIDE W IN POSTNATAL MOUSE HYPOTHALAMUS - A PUTATIVE REGULATOR OF ENERGY HOMEOSTASIS 査読

    T. Motoike, A. G. Skach, J. K. Godwin, C. M. Sinton, M. Yamazaki, M. Abe, R. Natsume, K. Sakimura, M. Yanagisawa

    NEUROSCIENCE   301   323 - 337   2015年8月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:PERGAMON-ELSEVIER SCIENCE LTD  

    Neuropeptide B and W (NPB and NPW) are cognate peptide ligands for NPBWR1 (GPR7), a G protein-coupled receptor. In rodents, they have been implicated in the regulation of energy homeostasis, neuroendocrine/autonomic responses, and social interactions. Although localization of these peptides and their receptors in adult rodent brain has been well documented, their expression in mouse brain during development is unknown. Here we demonstrate the transient expression of NPW mRNA in the dorsomedial hypothalamus (DMH) of postnatal mouse brain and its co-localization with neuropeptide Y (NPY) mRNA. Neurons expressing both NPW and NPY mRNAs begin to emerge in the DMH at about postnatal day 0 (P-0) through P-3. Their expression is highest around P-14, declines after P-21, and by P-28 only a faint expression of NPW and NPY mRNA remains. In P-18 brains, we detected NPW neurons in the region spanning the subincertal nucleus (SubI), the lateral hypothalamic (LH) perifornical (PF) areas, and the DMH, where the highest expression of NPW mRNA was observed. The majority of these postnatal hypothalamic NPW neurons co-express NPY mRNA. A cross of NPW-iCre knock-in mice with a Cre-dependent tdTomato reporter line revealed that more than half of the reporter-positive neurons in the adult DMH, which mature from the transiently NPW-expressing neurons, are sensitive to peripherally administrated leptin. These data suggest that the DMH neurons that transiently co-express NPW and NPY in the peri-weaning period might play a role in regulating energy homeostasis during postnatal development. (C) 2015 IBRO. Published by Elsevier Ltd. All rights reserved.

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  • Bcl11b SWI/SNF-complex subunit modulates intestinal adenoma and regeneration after γ-irradiation through Wnt/β-catenin pathway. 査読 国際誌

    Sakamaki A, Katsuragi Y, Otsuka K, Tomita M, Obata M, Iwasaki T, Abe M, Sato T, Ochiai M, Sakuraba Y, Aoyagi Y, Gondo Y, Sakimura K, Nakagama H, Mishima Y, Kominami R

    Carcinogenesis   36 ( 6 )   622 - 31   2015年6月

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    記述言語:英語  

    DOI: 10.1093/carcin/bgv044

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  • RASAL3, a novel hematopoietic RasGAP protein, regulates the number and functions of NKT cells. 査読 国際誌

    Suguru Saito, Toshihiko Kawamura, Masaya Higuchi, Takahiro Kobayashi, Manami Yoshita-Takahashi, Maya Yamazaki, Manabu Abe, Kenji Sakimura, Yasuhiro Kanda, Hiroki Kawamura, Shuying Jiang, Makoto Naito, Takumi Yoshizaki, Masahiko Takahashi, Masahiro Fujii

    European journal of immunology   45 ( 5 )   1512 - 23   2015年5月

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    記述言語:英語  

    Ras GTPase-activating proteins negatively regulate the Ras/Erk signaling pathway, thereby playing crucial roles in the proliferation, function, and development of various types of cells. In this study, we identified a novel Ras GTPase-activating proteins protein, RASAL3, which is predominantly expressed in cells of hematopoietic lineages, including NKT, B, and T cells. We established systemic RASAL3-deficient mice, and the mice exhibited a severe decrease in NKT cells in the liver at 8 weeks of age. The treatment of RASAL3-deficient mice with α-GalCer, a specific agonist for NKT cells, induced liver damage, but the level was less severe than that in RASAL3-competent mice, and the attenuated liver damage was accompanied by a reduced production of interleukin-4 and interferon-γ from NKT cells. RASAL3-deficient NKT cells treated with α-GalCer in vitro presented augmented Erk phosphorylation, suggesting that there is dysregulated Ras signaling in the NKT cells of RASAL3-deficient mice. Taken together, these results suggest that RASAL3 plays an important role in the expansion and functions of NKT cells in the liver by negatively regulating Ras/Erk signaling, and might be a therapeutic target for NKT-associated diseases.

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  • Bcl11b prevents the intrathymic development of innate CD8 T cells in a cell intrinsic manner. 査読 国際誌

    Satoshi Hirose, Maki Touma, Rieka Go, Yoshinori Katsuragi, Yoshiyuki Sakuraba, Yoichi Gondo, Manabu Abe, Kenji Sakimura, Yukio Mishima, Ryo Kominami

    International immunology   27 ( 4 )   205 - 15   2015年4月

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    記述言語:英語  

    If Bcl11b activity is compromised, CD4(+)CD8(+) double-positive (DP) thymocytes produce a greatly increased fraction of innate CD8(+) single-positive (SP) cells highly producing IFN-γ, which are also increased in mice deficient of genes such as Itk, Id3 and NF-κB1 that affect TCR signaling. Of interest, the increase in the former two is due to the bystander effect of IL-4 that is secreted by promyelocytic leukemia zinc finger-expressing NKT and γδT cells whereas the increase in the latter is cell intrinsic. Bcl11b zinc-finger proteins play key roles in T cell development and T cell-mediated immune response likely through TCR signaling. We examined thymocytes at and after the DP stage in Bcl11b (F/S826G) CD4cre, Bcl11b (F/+) CD4cre and Bcl11b (+/S826G) mice, carrying the allele that substituted serine for glycine at the position of 826. Here we show that Bcl11b impairment leads to an increase in the population of TCRαβ(high)CD44(high)CD122(high) innate CD8SP thymocytes, together with two different developmental abnormalities: impaired positive and negative selection accompanying a reduction in the number of CD8SP cells, and developmental arrest of NKT cells at multiple steps. The innate CD8SP thymocytes express Eomes and secrete IFN-γ after stimulation with PMA and ionomycin, and in this case their increase is not due to a bystander effect of IL-4 but cell intrinsic. Those results indicate that Bcl11b regulates development of different thymocyte subsets at multiple stages and prevents an excess of innate CD8SP thymocytes.

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  • Anterograde C1ql1 signaling is required in order to determine and maintain a single-winner climbing fiber in the mouse cerebellum. 査読 国際誌

    Wataru Kakegawa, Nikolaos Mitakidis, Eriko Miura, Manabu Abe, Keiko Matsuda, Yukari H Takeo, Kazuhisa Kohda, Junko Motohashi, Akiyo Takahashi, Soichi Nagao, Shin-ichi Muramatsu, Masahiko Watanabe, Kenji Sakimura, A Radu Aricescu, Michisuke Yuzaki

    Neuron   85 ( 2 )   316 - 29   2015年1月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Neuronal networks are dynamically modified by selective synapse pruning during development and adulthood. However, how certain connections win the competition with others and are subsequently maintained is not fully understood. Here, we show that C1ql1, a member of the C1q family of proteins, is provided by climbing fibers (CFs) and serves as a crucial anterograde signal to determine and maintain the single-winner CF in the mouse cerebellum throughout development and adulthood. C1ql1 specifically binds to the brain-specific angiogenesis inhibitor 3 (Bai3), which is a member of the cell-adhesion G-protein-coupled receptor family and expressed on postsynaptic Purkinje cells. C1ql1-Bai3 signaling is required for motor learning but not for gross motor performance or coordination. Because related family members of C1ql1 and Bai3 are expressed in various brain regions, the mechanism described here likely applies to synapse formation, maintenance, and function in multiple neuronal circuits essential for important brain functions.

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  • Heterozygous Disruption of Autism susceptibility candidate 2 Causes Impaired Emotional Control and Cognitive Memory. 査読 国際誌

    Kei Hori, Taku Nagai, Wei Shan, Asami Sakamoto, Manabu Abe, Maya Yamazaki, Kenji Sakimura, Kiyofumi Yamada, Mikio Hoshino

    PloS one   10 ( 12 )   e0145979   2015年

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    記述言語:英語  

    Mutations in the Autism susceptibility candidate 2 gene (AUTS2) have been associated with a broad range of psychiatric illnesses including autism spectrum disorders, intellectual disability and schizophrenia. We previously demonstrated that the cytoplasmic AUTS2 acts as an upstream factor for the Rho family small GTPase Rac1 and Cdc42 that regulate the cytoskeletal rearrangements in neural cells. Moreover, genetic ablation of the Auts2 gene in mice has resulted in defects in neuronal migration and neuritogenesis in the developing cerebral cortex caused by inactivation of Rac1-signaling pathway, suggesting that AUTS2 is required for neural development. In this study, we conducted a battery of behavioral analyses on Auts2 heterozygous mutant mice to examine the involvement of Auts2 in adult cognitive brain functions. Auts2-deficient mice displayed a decrease in exploratory behavior as well as lower anxiety-like behaviors in the absence of any motor dysfunction. Furthermore, the capability for novel object recognition and cued associative memory were impaired in Auts2 mutant mice. Social behavior and sensory motor gating functions were, however, normal in the mutant mice as assessed by the three-chamber test and prepulse inhibition test, respectively. Together, our findings indicate that AUTS2 is critical for the acquisition of neurocognitive function.

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  • Cytoskeletal regulation by AUTS2 in neuronal migration and neuritogenesis. 査読 国際誌

    Kei Hori, Taku Nagai, Wei Shan, Asami Sakamoto, Shinichiro Taya, Ryoya Hashimoto, Takashi Hayashi, Manabu Abe, Maya Yamazaki, Keiko Nakao, Tomoki Nishioka, Kenji Sakimura, Kiyofumi Yamada, Kozo Kaibuchi, Mikio Hoshino

    Cell reports   9 ( 6 )   2166 - 79   2014年12月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:CELL PRESS  

    Mutations in the Autism susceptibility candidate 2 gene (AUTS2), whose protein is believed to act in neuronal cell nuclei, have been associated with multiple psychiatric illnesses, including autism spectrum disorders, intellectual disability, and schizophrenia. Here we show that cytoplasmic AUTS2 is involved in the regulation of the cytoskeleton and neural development. Immunohistochemistry and fractionation studies show that AUTS2 localizes not only in nuclei, but also in the cytoplasm, including in the growth cones in the developing brain. AUTS2 activates Rac1 to induce lamellipodia but downregulates Cdc42 to suppress filopodia. Our loss-of-function and rescue experiments show that a cytoplasmic AUTS2-Rac1 pathway is involved in cortical neuronal migration and neuritogenesis in the developing brain. These findings suggest that cytoplasmic AUTS2 acts as a regulator of Rho family GTPases to contribute to brain development and give insight into the pathology of human psychiatric disorders with AUTS2 mutations.

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  • Proteasome dysfunction induces muscle growth defects and protein aggregation. 査読 国際誌

    Yasuo Kitajima, Yoshitaka Tashiro, Naoki Suzuki, Hitoshi Warita, Masaaki Kato, Maki Tateyama, Risa Ando, Rumiko Izumi, Maya Yamazaki, Manabu Abe, Kenji Sakimura, Hidefumi Ito, Makoto Urushitani, Ryoichi Nagatomi, Ryosuke Takahashi, Masashi Aoki

    Journal of cell science   127 ( Pt 24 )   5204 - 17   2014年12月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    The ubiquitin-proteasome and autophagy-lysosome pathways are the two major routes of protein and organelle clearance. The role of the proteasome pathway in mammalian muscle has not been examined in vivo. In this study, we report that the muscle-specific deletion of a crucial proteasomal gene, Rpt3 (also known as Psmc4), resulted in profound muscle growth defects and a decrease in force production in mice. Specifically, developing muscles in conditional Rpt3-knockout animals showed dysregulated proteasomal activity. The autophagy pathway was upregulated, but the process of autophagosome formation was impaired. A microscopic analysis revealed the accumulation of basophilic inclusions and disorganization of the sarcomeres in young adult mice. Our results suggest that appropriate proteasomal activity is important for muscle growth and for maintaining myofiber integrity in collaboration with autophagy pathways. The deletion of a component of the proteasome complex contributed to myofiber degeneration and weakness in muscle disorders that are characterized by the accumulation of abnormal inclusions.

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  • IL1RAPL1 knockout mice show spine density decrease, learning deficiency, hyperactivity and reduced anxiety-like behaviours. 査読 国際誌

    Misato Yasumura, Tomoyuki Yoshida, Maya Yamazaki, Manabu Abe, Rie Natsume, Kouta Kanno, Takeshi Uemura, Keizo Takao, Kenji Sakimura, Takefumi Kikusui, Tsuyoshi Miyakawa, Masayoshi Mishina

    Scientific reports   4   6613 - 6613   2014年10月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:NATURE PUBLISHING GROUP  

    IL-1 receptor accessory protein-like 1 (IL1RAPL1) is responsible for nonsyndromic intellectual disability and is associated with autism. IL1RAPL1 mediates excitatory synapse formation through trans-synaptic interaction with PTPδ. Here, we showed that the spine density of cortical neurons was significantly reduced in IL1RAPL1 knockout mice. The spatial reference and working memories and remote fear memory were mildly impaired in IL1RAPL1 knockout mice. Furthermore, the behavioural flexibility was slightly reduced in the T-maze test. Interestingly, the performance of IL1RAPL1 knockout mice in the rotarod test was significantly better than that of wild-type mice. Moreover, IL1RAPL1 knockout mice consistently exhibited high locomotor activity in all the tasks examined. In addition, open-space and height anxiety-like behaviours were decreased in IL1RAPL1 knockout mice. These results suggest that IL1RAPL1 ablation resulted in spine density decrease and affected not only learning but also behavioural flexibility, locomotor activity and anxiety.

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  • Visualization of corticotropin-releasing factor neurons by fluorescent proteins in the mouse brain and characterization of labeled neurons in the paraventricular nucleus of the hypothalamus. 査読 国際誌

    Keiichi Itoi, Ashraf Hossain Talukder, Toshimitsu Fuse, Takuji Kaneko, Ryo Ozawa, Takayuki Sato, Takuma Sugaya, Katsuya Uchida, Maya Yamazaki, Manabu Abe, Rie Natsume, Kenji Sakimura

    Endocrinology   155 ( 10 )   4054 - 60   2014年10月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ENDOCRINE SOC  

    Corticotropin-releasing factor (CRF) is the key regulator of the hypothalamic-pituitary-adrenal axis. CRF neurons cannot be distinguished morphologically from other neuroendocrine neurons in the paraventricular nucleus of the hypothalamus (PVH) without immunostaining. Thus, we generated a knock-in mouse that expresses modified yellow fluorescent protein (Venus) in CRF neurons (CRF-Venus), and yet its expression is driven by the CRF promoter and responds to changes in the interior milieu. In CRF-Venus, Venus-expressing neurons were distributed in brain regions harboring CRF neurons, including the PVH. The majority of Venus-expressing neurons overlapped with CRF-expressing neurons in the PVH, but many neurons expressed only Venus or CRF in a physiological glucocorticoid condition. After glucocorticoid deprivation, however, Venus expression intensified, and most Venus neurons coexpressed CRF. Conversely, Venus expression was suppressed by excess glucocorticoids. Expression of copeptin, a peptide encoded within the vasopressin gene, was induced in PVH-Venus neurons by glucocorticoid deprivation and suppressed by glucocorticoid administration. Thus, Venus neurons recapitulated glucocorticoid-dependent vasopressin expression in PVH-CRF neurons. Noradrenaline increased the frequency of glutamate-dependent excitatory postsynaptic currents recorded from Venus-expressing neurons in the voltage clamp mode. In addition, the CRF-iCre knock-in mouse was crossed with a CAG-CAT-EGFP reporter mouse to yield the Tg(CAG-CAT-EGFP/wt);CRF(iCre/wt) (EGFP/CRF-iCre) mouse, in which enhanced green fluorescent protein (EGFP) is driven by the CAG promoter. EGFP was expressed more constitutively in the PVH of EGFP/CRF-iCre mice. Thus, CRF-Venus may have an advantage for monitoring dynamic changes in CRF neurons and CRF networks in different glucocorticoid states.

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  • Cdk5/p35 is required for motor coordination and cerebellar plasticity. 査読 国際誌

    Xiaojuan He, Masato Ishizeki, Naoki Mita, Seitaro Wada, Yoshifumi Araki, Hiroo Ogura, Manabu Abe, Maya Yamazaki, Kenji Sakimura, Katsuhiko Mikoshiba, Takafumi Inoue, Toshio Ohshima

    Journal of neurochemistry   131 ( 1 )   53 - 64   2014年10月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:WILEY-BLACKWELL  

    Previous studies have implicated the role of Purkinje cells in motor learning and the underlying mechanisms have also been identified in great detail during the last decades. Here we report that cyclin-dependent kinase 5 (Cdk5)/p35 in Purkinje cell also contributes to synaptic plasticity. We previously showed that p35(-/-) (p35 KO) mice exhibited a subtle abnormality in brain structure and impaired spatial learning and memory. Further behavioral analysis showed that p35 KO mice had a motor coordination defect, suggesting that p35, one of the activators of Cdk5, together with Cdk5 may play an important role in cerebellar motor learning. Therefore, we created Purkinje cell-specific conditional Cdk5/p35 knockout (L7-p35 cKO) mice, analyzed the cerebellar histology and Purkinje cell morphology of these mice, evaluated their performance with balance beam and rota-rod test, and performed electrophysiological recordings to assess long-term synaptic plasticity. Our analyses showed that Purkinje cell-specific deletion of Cdk5/p35 resulted in no changes in Purkinje cell morphology but severely impaired motor coordination. Furthermore, disrupted cerebellar long-term synaptic plasticity was observed at the parallel fiber-Purkinje cell synapse in L7-p35 cKO mice. These results indicate that Cdk5/p35 is required for motor learning and involved in long-term synaptic plasticity.

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  • Expansion of stochastic expression repertoire by tandem duplication in mouse Protocadherin-α cluster. 査読 国際誌

    Kaneko R, Abe M, Hirabayashi T, Uchimura A, Sakimura K, Yanagawa Y, Yagi T

    Scientific reports   4   6263 - 6263   2014年9月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

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  • Detailed expression pattern of aldolase C (Aldoc) in the cerebellum, retina and other areas of the CNS studied in Aldoc-Venus knock-in mice. 査読 国際誌

    Hirofumi Fujita, Hanako Aoki, Itsuki Ajioka, Maya Yamazaki, Manabu Abe, Arata Oh-Nishi, Kenji Sakimura, Izumi Sugihara

    PloS one   9 ( 1 )   e86679   2014年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:PUBLIC LIBRARY SCIENCE  

    Aldolase C (Aldoc, also known as "zebrin II"), a brain type isozyme of a glycolysis enzyme, is expressed heterogeneously in subpopulations of cerebellar Purkinje cells (PCs) that are arranged longitudinally in a complex striped pattern in the cerebellar cortex, a pattern which is closely related to the topography of input and output axonal projections. Here, we generated knock-in Aldoc-Venus mice in which Aldoc expression is visualized by expression of a fluorescent protein, Venus. Since there was no obvious phenotypes in general brain morphology and in the striped pattern of the cerebellum in mutants, we made detailed observation of Aldoc expression pattern in the nervous system by using Venus expression in Aldoc-Venus heterozygotes. High levels of Venus expression were observed in cerebellar PCs, cartwheel cells in the dorsal cochlear nucleus, sensory epithelium of the inner ear and in all major types of retinal cells, while moderate levels of Venus expression were observed in astrocytes and satellite cells in the dorsal root ganglion. The striped arrangement of PCs that express Venus to different degrees was carefully traced with serial section alignment analysis and mapped on the unfolded scheme of the entire cerebellar cortex to re-identify all individual Aldoc stripes. A longitudinally striped boundary of Aldoc expression was first identified in the mouse flocculus, and was correlated with the climbing fiber projection pattern and expression of another compartmental marker molecule, heat shock protein 25 (HSP25). As in the rat, the cerebellar nuclei were divided into the rostrodorsal negative and the caudoventral positive portions by distinct projections of Aldoc-positive and negative PC axons in the mouse. Identification of the cerebellar Aldoc stripes in this study, as indicated in sample coronal and horizontal sections as well as in sample surface photos of whole-mount preparations, can be referred to in future experiments.

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  • Point mutation in syntaxin-1A causes abnormal vesicle recycling, behaviors, and short term plasticity. 査読 国際誌

    Yumi Watanabe, Norikazu Katayama, Kosei Takeuchi, Tetsuya Togano, Rieko Itoh, Michiko Sato, Maya Yamazaki, Manabu Abe, Toshiya Sato, Kanako Oda, Minesuke Yokoyama, Keizo Takao, Masahiro Fukaya, Tsuyoshi Miyakawa, Masahiko Watanabe, Kenji Sakimura, Toshiya Manabe, Michihiro Igarashi

    The Journal of biological chemistry   288 ( 48 )   34906 - 19   2013年11月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Syntaxin-1A is a t-SNARE that is involved in vesicle docking and vesicle fusion; it is important in presynaptic exocytosis in neurons because it interacts with many regulatory proteins. Previously, we found the following: 1) that autophosphorylated Ca(2+)/calmodulin-dependent protein kinase II (CaMKII), an important modulator of neural plasticity, interacts with syntaxin-1A to regulate exocytosis, and 2) that a syntaxin missense mutation (R151G) attenuated this interaction. To determine more precisely the physiological importance of this interaction between CaMKII and syntaxin, we generated mice with a knock-in (KI) syntaxin-1A (R151G) mutation. Complexin is a molecular clamp involved in exocytosis, and in the KI mice, recruitment of complexin to the SNARE complex was reduced because of an abnormal CaMKII/syntaxin interaction. Nevertheless, SNARE complex formation was not inhibited, and consequently, basal neurotransmission was normal. However, the KI mice did exhibit more enhanced presynaptic plasticity than wild-type littermates; this enhanced plasticity could be associated with synaptic response than did wild-type littermates; this pronounced response included several behavioral abnormalities. Notably, the R151G phenotypes were generally similar to previously reported CaMKII mutant phenotypes. Additionally, synaptic recycling in these KI mice was delayed, and the density of synaptic vesicles was reduced. Taken together, our results indicated that this single point mutation in syntaxin-1A causes abnormal regulation of neuronal plasticity and vesicle recycling and that the affected syntaxin-1A/CaMKII interaction is essential for normal brain and synaptic functions in vivo.

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  • Cell of origin in radiation-induced premalignant thymocytes with differentiation capability in mice conditionally losing one Bcl11b allele

    Rieka Go, Satoshi Hirose, Yoshinori Katsuragi, Miki Obata, Manabu Abe, Yukio Mishima, Kenji Sakimura, Ryo Kominami

    Cancer Science   104 ( 8 )   1009 - 1016   2013年8月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Bcl11b is a haploinsufficient tumor suppressor, mutations or deletion of which has been found in 10-16% of T-cell acute lymphoblastic leukemias. Bcl11bKO/+ heterozygous mice are susceptible to thymic lymphomas, a model of T-cell acute lymphoblastic leukemia, when γ-irradiated, and irradiated Bcl11bKO/+ mice generate clonally expanding or premalignant thymocytes before thymic lymphoma development. Cells with radiation-induced DNA damages are assumed to be the cells of origin in tumors
    however, which thymocyte is the tumor cell origin remains obscure. In this study we generated Bcl11bflox/+
    Lck-Cre and Bcl11bflox/+
    CD4-Cre mice
    in the former, loss of one Bcl11b allele occurs in thymocytes at the immature CD4-CD8- stage, whereas in the latter the loss occurs in the more differentiated CD4+CD8+ double-positive stage. We examined clonal expansion and differentiation of thymocytes in mice 60 days after 3 Gy γ-irradiation. Half (9/18) of the thymuses in the Bcl11bflox/+
    Lck-Cre group showed limited rearrangement sites at the T-cell receptor-β (TCRβ) locus, indicating clonal cell expansion, but none in the Bcl11bflox/+
    CD4-Cre group did. This indicates that the origin of the premalignant thymocytes is not in double-positive cells but immature thymocytes. Interestingly, those premalignant thymocytes underwent rearrangement at various different sites of the TCRα locus and the majority showed a higher expression of TCRβ and CD8, and more differentiated phenotypes. This suggests the existence of a subpopulation of immature cells within the premalignant cells that is capable of proliferating and continuously producing differentiated thymocytes. © 2013 Japanese Cancer Association.

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  • A critical role of serotonergic inhibitory projections to orexin neurons in regulation of sleep/wakefulness states 査読

    Natsuko Tsujino, Yuki Saito, Mari Hondo, Manabu Abe, Kenji Sakimura, Takeshi Sakurai

    JOURNAL OF PHYSIOLOGICAL SCIENCES   63 ( 1 )   S147 - S147   2013年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:SPRINGER JAPAN KK  

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  • GABAergic neurons in the preoptic area send direct inhibitory projections to orexin neurons. 査読 国際誌

    Yuki C Saito, Natsuko Tsujino, Emi Hasegawa, Kaori Akashi, Manabu Abe, Michihiro Mieda, Kenji Sakimura, Takeshi Sakurai

    Frontiers in neural circuits   7   192 - 192   2013年

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    記述言語:英語  

    Populations of neurons in the hypothalamic preoptic area (POA) fire rapidly during sleep, exhibiting sleep/waking state-dependent firing patterns that are the reciprocal of those observed in the arousal system. The majority of these preoptic "sleep-active" neurons contain the inhibitory neurotransmitter GABA. On the other hand, a population of neurons in the lateral hypothalamic area (LHA) contains orexins, which play an important role in the maintenance of wakefulness, and exhibit an excitatory influence on arousal-related neurons. It is important to know the anatomical and functional interactions between the POA sleep-active neurons and orexin neurons, both of which play important, but opposite roles in regulation of sleep/wakefulness states. In this study, we confirmed that specific pharmacogenetic stimulation of GABAergic neurons in the POA leads to an increase in the amount of non-rapid eye movement (NREM) sleep. We next examined direct connectivity between POA GABAergic neurons and orexin neurons using channelrhodopsin 2 (ChR2) as an anterograde tracer as well as an optogenetic tool. We expressed ChR2-eYFP selectively in GABAergic neurons in the POA by AAV-mediated gene transfer, and examined the projection sites of ChR2-eYFP-expressing axons, and the effect of optogenetic stimulation of ChR2-eYFP on the activity of orexin neurons. We found that these neurons send widespread projections to wakefulness-related areas in the hypothalamus and brain stem, including the LHA where these fibers make close appositions to orexin neurons. Optogenetic stimulation of these fibers resulted in rapid inhibition of orexin neurons. These observations suggest direct connectivity between POA GABAergic neurons and orexin neurons.

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  • Motor neuron-specific disruption of proteasomes, but not autophagy, replicates amyotrophic lateral sclerosis. 査読 国際誌

    Yoshitaka Tashiro, Makoto Urushitani, Haruhisa Inoue, Masato Koike, Yasuo Uchiyama, Masaaki Komatsu, Keiji Tanaka, Maya Yamazaki, Manabu Abe, Hidemi Misawa, Kenji Sakimura, Hidefumi Ito, Ryosuke Takahashi

    The Journal of biological chemistry   287 ( 51 )   42984 - 94   2012年12月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Evidence suggests that protein misfolding is crucially involved in the pathogenesis of amyotrophic lateral sclerosis (ALS). However, controversy still exists regarding the involvement of proteasomes or autophagy in ALS due to previous conflicting results. Here, we show that impairment of the ubiquitin-proteasome system, but not the autophagy-lysosome system in motor neurons replicates ALS in mice. Conditional knock-out mice of the proteasome subunit Rpt3 in a motor neuron-specific manner (Rpt3-CKO) showed locomotor dysfunction accompanied by progressive motor neuron loss and gliosis. Moreover, diverse ALS-linked proteins, including TAR DNA-binding protein 43 kDa (TDP-43), fused in sarcoma (FUS), ubiquilin 2, and optineurin were mislocalized or accumulated in motor neurons, together with other typical ALS hallmarks such as basophilic inclusion bodies. On the other hand, motor neuron-specific knock-out of Atg7, a crucial component for the induction of autophagy (Atg7-CKO), only resulted in cytosolic accumulation of ubiquitin and p62, and no TDP-43 or FUS pathologies or motor dysfunction was observed. These results strongly suggest that proteasomes, but not autophagy, fundamentally govern the development of ALS in which TDP-43 and FUS proteinopathy may play a crucial role. Enhancement of proteasome activity may be a promising strategy for the treatment of ALS.

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  • Synapse type-independent degradation of the endocannabinoid 2-arachidonoylglycerol after retrograde synaptic suppression. 査読 国際誌

    Asami Tanimura, Motokazu Uchigashima, Maya Yamazaki, Naofumi Uesaka, Takayasu Mikuni, Manabu Abe, Kouichi Hashimoto, Masahiko Watanabe, Kenji Sakimura, Masanobu Kano

    Proceedings of the National Academy of Sciences of the United States of America   109 ( 30 )   12195 - 200   2012年7月

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    記述言語:英語  

    The endocannabinoid 2-arachidonoylglycerol (2-AG) mediates retrograde synaptic suppression. Although the mechanisms of 2-AG production are well characterized, how 2-AG is degraded is less clearly understood. Here we found that expression of the 2-AG hydrolyzing enzyme monoacylglycerol lipase (MGL) was highly heterogeneous in the cerebellum, being rich within parallel fiber (PF) terminals, weak in Bergman glia (BG), and absent in other synaptic terminals. Despite this highly selective MGL expression pattern, 2-AG-mediated retrograde suppression was significantly prolonged at not only PF-Purkinje cell (PC) synapses but also climbing fiber-PC synapses in granule cell-specific MGL knockout (MGL-KO) mice whose cerebellar MGL expression was confined to the BG. Virus-mediated expression of MGL into the BG of global MGL-KO mice significantly shortened 2-AG-mediated retrograde suppression at PF-PC synapses. Furthermore, contribution of MGL to termination of 2-AG signaling depended on the distance from MGL-rich PFs to inhibitory synaptic terminals. Thus, 2-AG is degraded in a synapse-type independent manner by MGL present in PFs and the BG. The results of the present study strongly suggest that MGL regulates 2-AG signaling rather broadly within a certain range of neural tissue, although MGL expression is heterogeneous and limited to a subset of nerve terminals and astrocytes.

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  • Inverse synaptic tagging of inactive synapses via dynamic interaction of Arc/Arg3.1 with CaMKIIβ. 査読 国際誌

    Okuno H, Akashi K, Ishii Y, Yagishita-Kyo N, Suzuki K, Nonaka M, Kawashima T, Fujii H, Takemoto-Kimura S, Abe M, Natsume R, Chowdhury S, Sakimura K, Worley PF, Bito H

    Cell   149 ( 4 )   886 - 98   2012年5月

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  • Cav2.1 in cerebellar Purkinje cells regulates competitive excitatory synaptic wiring, cell survival, and cerebellar biochemical compartmentalization. 査読 国際誌

    Taisuke Miyazaki, Miwako Yamasaki, Kouichi Hashimoto, Maya Yamazaki, Manabu Abe, Hiroshi Usui, Masanobu Kano, Kenji Sakimura, Masahiko Watanabe

    The Journal of neuroscience : the official journal of the Society for Neuroscience   32 ( 4 )   1311 - 28   2012年1月

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    記述言語:英語  

    In the adult cerebellum, each Purkinje cell (PC) is innervated by a single climbing fiber (CF) in proximal dendrites and 10(5)-10(6) parallel fibers (PFs) in distal dendrites. This organized wiring is established postnatally through heterosynaptic competition between PFs and CFs and homosynaptic competition among multiple CFs. Using PC-specific Cav2.1 knock-out mice (PC-Cav2.1 KO mice), we have demonstrated recently that postsynaptic Cav2.1 plays a key role in the homosynaptic competition by promoting functional strengthening and dendritic translocation of single "winner" CFs. Here, we report that Cav2.1 in PCs, but not in granule cells, is also essential for the heterosynaptic competition. In PC-Cav2.1 KO mice, the extent of CF territory was limited to the soma and basal dendrites, whereas PF territory was expanded reciprocally. Consequently, the proximal somatodendritic domain of PCs displayed hyperspiny transformation and fell into chaotic innervation by multiple CFs and numerous PFs. PC-Cav2.1 KO mice also displayed patterned degeneration of PCs, which occurred preferentially in aldolase C/zebrin II-negative cerebellar compartments. Furthermore, the mutually complementary expression of phospholipase Cβ3 (PLCβ3) and PLCβ4 was altered such that their normally sharp boundary was blurred in the PCs of PC-Cav2.1 KO mice. This blurring was caused by an impaired posttranscriptional downregulation of PLCβ3 in PLCβ4-dominant PCs during the early postnatal period. A similar alteration was noted in the banded expression of the glutamate transporter EAAT4 in PC-Cav2.1 KO mice. Therefore, Cav2.1 in PCs is essential for competitive synaptic wiring, cell survival, and the establishment of precise boundaries and reciprocity of biochemical compartments in PCs.

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  • Basic leucine zipper transcription factor, ATF-like (BATF) regulates epigenetically and energetically effector CD8 T-cell differentiation via Sirt1 expression. 査読 国際誌

    Shoko Kuroda, Maya Yamazaki, Manabu Abe, Kenji Sakimura, Hiroshi Takayanagi, Yoshiko Iwai

    Proceedings of the National Academy of Sciences of the United States of America   108 ( 36 )   14885 - 9   2011年9月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:NATL ACAD SCIENCES  

    CD8 T cells play a critical role in protection against viral infections. During effector differentiation, CD8 T cells dramatically change chromatin structure and cellular metabolism, but how energy production increases in response to these epigenetic changes is unknown. We found that loss of basic leucine zipper transcription factor, ATF-like (BATF) inhibited effector CD8 T-cell differentiation. At the late effector stage, BATF was induced by IL-12 and required for IL-12-mediated histone acetylation and survival of effector T cells. BATF, together with c-Jun, transcriptionally inhibited expression of the nicotinamide adenine dinucleotide (NAD(+))-dependent deacetylase Sirt1, resulting in increased histone acetylation of the T-bet locus and increased cellular NAD(+), which increased ATP production. In turn, high levels of T-bet expression and ATP production promoted effector differentiation and cell survival. These results suggest that BATF promotes effector CD8 T-cell differentiation by regulating both epigenetic remodeling and energy metabolism through Sirt1 expression.

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  • Glutamate receptor δ2 is essential for input pathway-dependent regulation of synaptic AMPAR contents in cerebellar Purkinje cells. 査読 国際誌

    Yamasaki M, Miyazaki T, Azechi H, Abe M, Natsume R, Hagiwara T, Aiba A, Mishina M, Sakimura K, Watanabe M

    The Journal of neuroscience : the official journal of the Society for Neuroscience   31 ( 9 )   3362 - 74   2011年3月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

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  • LPA4 regulates blood and lymphatic vessel formation during mouse embryogenesis. 査読 国際誌

    Hayakazu Sumida, Kyoko Noguchi, Yasuyuki Kihara, Manabu Abe, Keisuke Yanagida, Fumie Hamano, Shinichi Sato, Kunihiko Tamaki, Yasuyuki Morishita, Mitsunobu R Kano, Caname Iwata, Kohei Miyazono, Kenji Sakimura, Takao Shimizu, Satoshi Ishii

    Blood   116 ( 23 )   5060 - 70   2010年12月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Lysophosphatidic acid (LPA) is a potent lipid mediator with a wide variety of biological actions mediated through G protein-coupled receptors (LPA(1-6)). LPA(4) has been identified as a G(13) protein-coupled receptor, but its physiological role is unknown. Here we show that a subset of LPA(4)-deficient embryos did not survive gestation and displayed hemorrhages and/or edema in many organs at multiple embryonic stages. The blood vessels of bleeding LPA(4)-deficient embryos were often dilated. The recruitment of mural cells, namely smooth muscle cells and pericytes, was impaired. Consistently, Matrigel plug assays showed decreased mural cell coverage of endothelial cells in the neovessels of LPA(4)-deficient adult mice. In situ hybridization detected Lpa4 mRNA in the endothelium of some vasculatures. Similarly, the lymphatic vessels of edematous embryos were dilated. These results suggest that LPA(4) regulates establishment of the structure and function of blood and lymphatic vessels during mouse embryogenesis. Considering the critical role of autotaxin (an enzyme involved in LPA production) and Gα(13) in vascular development, we suggest that LPA(4) provides a link between these 2 molecules.

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  • Chondroitin sulfate N-acetylgalactosaminyltransferase-1 is required for normal cartilage development. 査読 国際誌

    Yumi Watanabe, Kosei Takeuchi, Susumu Higa Onaga, Michiko Sato, Mika Tsujita, Manabu Abe, Rie Natsume, Minqi Li, Tatsuya Furuichi, Mika Saeki, Tomomi Izumikawa, Ayumi Hasegawa, Minesuke Yokoyama, Shiro Ikegawa, Kenji Sakimura, Norio Amizuka, Hiroshi Kitagawa, Michihiro Igarashi

    The Biochemical journal   432 ( 1 )   47 - 55   2010年11月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    CS (chondroitin sulfate) is a glycosaminoglycan species that is widely distributed in the extracellular matrix. To understand the physiological roles of enzymes involved in CS synthesis, we produced CSGalNAcT1 (CS N-acetylgalactosaminyltransferase 1)-null mice. CS production was reduced by approximately half in CSGalNAcT1-null mice, and the amount of short-chain CS was also reduced. Moreover, the cartilage of the null mice was significantly smaller than that of wild-type mice. Additionally, type-II collagen fibres in developing cartilage were abnormally aggregated and disarranged in the homozygous mutant mice. These results suggest that CSGalNAcT1 is required for normal CS production in developing cartilage.

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  • Calpain 8/nCL-2 and calpain 9/nCL-4 constitute an active protease complex, G-calpain, involved in gastric mucosal defense. 査読 国際誌

    Shoji Hata, Manabu Abe, Hidenori Suzuki, Fujiko Kitamura, Noriko Toyama-Sorimachi, Keiko Abe, Kenji Sakimura, Hiroyuki Sorimachi

    PLoS genetics   6 ( 7 )   e1001040   2010年7月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Calpains constitute a superfamily of Ca2+-dependent cysteine proteases, indispensable for various cellular processes. Among the 15 mammalian calpains, calpain 8/nCL-2 and calpain 9/nCL-4 are predominantly expressed in the gastrointestinal tract and are restricted to the gastric surface mucus (pit) cells in the stomach. Possible functions reported for calpain 8 are in vesicle trafficking between ER and Golgi, and calpain 9 are implicated in suppressing tumorigenesis. These highlight that calpains 8 and 9 are regulated differently from each other and from conventional calpains and, thus, have potentially important, specific functions in the gastrointestinal tract. However, there is no direct evidence implicating calpain 8 or 9 in human disease, and their properties and physiological functions are currently unknown. To address their physiological roles, we analyzed mice with mutations in the genes for these calpains, Capn8 and Capn9. Capn8(-/-) and Capn9(-/-) mice were fertile, and their gastric mucosae appeared normal. However, both mice were susceptible to gastric mucosal injury induced by ethanol administration. Moreover, the Capn8(-/-) stomach showed significant decreases in both calpains 9 and 8, and the same was true for Capn9(-/-). Consistent with this finding, in the wild-type stomach, calpains 8 and 9 formed a complex we termed "G-calpain," in which both were essential for activity. This is the first example of a "hybrid" calpain complex. To address the physiological relevance of the calpain 8 proteolytic activity, we generated calpain 8:C105S "knock-in" (Capn8(CS/CS)) mice, which expressed a proteolytically inactive, but structurally intact, calpain 8. Although, unlike the Capn8(-/-) stomach, that of the Capn8(CS/CS) mice expressed a stable and active calpain 9, the mice were susceptible to ethanol-induced gastric injury. These results provide the first evidence that both of the gastrointestinal-tract-specific calpains are essential for gastric mucosal defense, and they point to G-calpain as a potential target for gastropathies caused by external stresses.

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  • TARPs gamma-2 and gamma-7 are essential for AMPA receptor expression in the cerebellum. 査読 国際誌

    Maya Yamazaki, Masahiro Fukaya, Kouichi Hashimoto, Miwako Yamasaki, Mika Tsujita, Makoto Itakura, Manabu Abe, Rie Natsume, Masami Takahashi, Masanobu Kano, Kenji Sakimura, Masahiko Watanabe

    The European journal of neuroscience   31 ( 12 )   2204 - 20   2010年6月

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    記述言語:英語  

    The alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA)-type glutamate receptors require auxiliary subunits termed transmembrane AMPA receptor regulatory proteins (TARPs), which promote receptor trafficking to the cell surface and synapses and modulate channel pharmacology and gating. Of six TARPs, gamma-2 and gamma-7 are the two major TARPs expressed in the cerebellum. In the present study, we pursued their roles in synaptic expression of cerebellar AMPA receptors. In the cerebellar cortex, gamma-2 and gamma-7 were preferentially localized at various asymmetrical synapses. Using quantitative Western blot and immunofluorescence, we found severe reductions in GluA2 and GluA3 and mild reduction in GluA4 in gamma-2-knockout (KO) cerebellum, whereas GluA1 and GluA4 were moderately reduced in gamma-7-KO cerebellum. GluA2, GluA3 and GluA4 were further reduced in gamma-2/gamma-7 double-KO (DKO) cerebellum. The large losses of GluA2 and GluA3 in gamma-2-KO mice and further reductions in DKO mice were confirmed at all asymmetrical synapses examined with postembedding immunogold. Most notably, the GluA2 level in the postsynaptic density fraction, GluA2 labeling density at parallel fiber-Purkinje cell synapses, and AMPA receptor-mediated currents at climbing fiber-Purkinje cell synapses were all reduced to approximately 10% of the wild-type levels in DKO mice. On the other hand, the reduction in GluA4 in gamma-7-KO granular layer reflected its loss at mossy fiber-granule cell synapses, whereas that of GluA1 and GluA4 in gamma-7-KO molecular layer was caused, at least partly, by their loss in Bergmann glia. Therefore, gamma-2 and gamma-7 cooperatively promote synaptic expression of cerebellar AMPA receptors, and the latter also promotes glial expression.

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  • The endocannabinoid 2-arachidonoylglycerol produced by diacylglycerol lipase alpha mediates retrograde suppression of synaptic transmission. 査読 国際誌

    Asami Tanimura, Maya Yamazaki, Yuki Hashimotodani, Motokazu Uchigashima, Shinya Kawata, Manabu Abe, Yoshihiro Kita, Kouichi Hashimoto, Takao Shimizu, Masahiko Watanabe, Kenji Sakimura, Masanobu Kano

    Neuron   65 ( 3 )   320 - 7   2010年2月

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    記述言語:英語  

    Endocannabinoids are released from postsynaptic neurons and cause retrograde suppression of synaptic transmission. Anandamide and 2-arachidonoylglycerol (2-AG) are regarded as two major endocannabinoids. To determine to what extent 2-AG contributes to retrograde signaling, we generated and analyzed mutant mice lacking either of the two 2-AG synthesizing enzymes diacylglycerol lipase alpha (DGLalpha) and beta (DGLbeta). We found that endocannabinoid-mediated retrograde synaptic suppression was totally absent in the cerebellum, hippocampus, and striatum of DGLalpha knockout mice, whereas the retrograde suppression was intact in DGLbeta knockout brains. The basal 2-AG content was markedly reduced and stimulus-induced elevation of 2-AG was absent in DGLalpha knockout brains, whereas the 2-AG content was normal in DGLbeta knockout brains. Morphology of the brain and expression of molecules required for 2-AG production other than DGLs were normal in the two knockout mice. We conclude that 2-AG produced by DGLalpha, but not by DGLbeta, mediates retrograde suppression at central synapses.

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  • NMDA receptor GluN2B (GluR epsilon 2/NR2B) subunit is crucial for channel function, postsynaptic macromolecular organization, and actin cytoskeleton at hippocampal CA3 synapses. 査読 国際誌

    Kaori Akashi, Toshikazu Kakizaki, Haruyuki Kamiya, Masahiro Fukaya, Miwako Yamasaki, Manabu Abe, Rie Natsume, Masahiko Watanabe, Kenji Sakimura

    The Journal of neuroscience : the official journal of the Society for Neuroscience   29 ( 35 )   10869 - 82   2009年9月

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    記述言語:英語  

    GluN2B (GluRepsilon2/NR2B) subunit is involved in synapse development, synaptic plasticity, and cognitive function. However, its roles in synaptic expression and function of NMDA receptors (NMDARs) in the brain remain mostly unknown because of the neonatal lethality of global knock-out mice. To address this, we generated conditional knock-out mice, in which GluN2B was ablated exclusively in hippocampal CA3 pyramidal cells. By immunohistochemistry, GluN2B disappeared and GluN1 (GluRzeta1/NR1) was moderately reduced, whereas GluN2A (GluRepsilon1/NR2A) and postsynaptic density-95 (PSD-95) were unaltered in the mutant CA3. This was consistent with protein contents in the CA3 crude fraction: 9.6% of control level for GluN2B, 47.7% for GluN1, 90.6% for GluN2A, and 98.0% for PSD-95. Despite the remaining NMDARs, NMDAR-mediated currents and long-term potentiation were virtually lost at various CA3 synapses. Then, we compared synaptic NMDARs by postembedding immunogold electron microscopy and immunoblot using the PSD fraction. In the mutant CA3, GluN1 was severely reduced in both immunogold (20.6-23.6%) and immunoblot (24.6%), whereas GluN2A and PSD-95 were unchanged in immunogold but markedly reduced in the PSD fraction (51.4 and 36.5%, respectively), indicating increased detergent solubility of PSD molecules. No such increased solubility was observed for GluN2B in the CA3 of GluN2A-knock-out mice. Furthermore, significant decreases were found in the ratio of filamentous to globular actin (49.5%) and in the density of dendritic spines (76.2%). These findings suggest that GluN2B is critically involved in NMDAR channel function, organization of postsynaptic macromolecular complexes, formation or maintenance of dendritic spines, and regulation of the actin cytoskeleton.

    DOI: 10.1523/JNEUROSCI.5531-08.2009

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  • Cysteinyl leukotriene 2 receptor-mediated vascular permeability via transendothelial vesicle transport. 査読 国際誌

    Michael P W Moos, Jeffrey D Mewburn, Frederick W K Kan, Satoshi Ishii, Manabu Abe, Kenji Sakimura, Kyoko Noguchi, Takao Shimizu, Colin D Funk

    FASEB journal : official publication of the Federation of American Societies for Experimental Biology   22 ( 12 )   4352 - 62   2008年12月

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    記述言語:英語  

    Cysteinyl leukotrienes (CysLTs) are potent mediators of inflammation synthesized by the concerted actions of 5-lipoxygenase (5-LO), 5-LO-activating protein (FLAP), leukotriene C(4) synthase, and additional downstream enzymes, starting with arachidonic acid substrate. CysLTs produced by macrophages, eosinophils, mast cells, and other inflammatory cells activate 3 different high-affinity CysLT receptors: CysLT(1)R, CysLT(2)R, and GPR 17. We sought to investigate vascular sites of CysLT(2)R expression and the role and mechanism of this receptor in mediating vascular permeability events. Vascular expression of CysLT(2)R was investigated by reporter gene expression in a novel CysLT(2)R deficient-LacZ mouse model. CysLT(2)R was expressed in small, but not large, vessels in mouse brain, bladder, skin, and cremaster muscle. Intravital, in addition to confocal and electron, microscopy investigations using FITC-labeled albumin in cremaster postcapillary venule preparations indicated rapid CysLT-mediated permeability, which was blocked by application of BAY-u9773, a dual CysLT(1)R/CysLT(2)R antagonist or by CysLT(2)R deficiency. Endothelial human CysLT(2)R overexpression in mice exacerbated vascular leakage even in the absence of exogenous ligand. The enhanced vascular permeability mediated by CysLT(2)R takes place via a transendothelial vesicle transport mechanism as opposed to a paracellular route and is controlled via Ca(2+) signaling. Our results reveal that CysLT(2)R can mediate inflammatory reactions in a vascular bed-specific manner by altering transendothelial vesicle transport-based vascular permeability.

    DOI: 10.1096/fj.08-113274

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  • Serine racemase is predominantly localized in neurons in mouse brain. 査読 国際誌

    Kazushi Miya, Ran Inoue, Yoshimi Takata, Manabu Abe, Rie Natsume, Kenji Sakimura, Kazuhisa Hongou, Toshio Miyawaki, Hisashi Mori

    The Journal of comparative neurology   510 ( 6 )   641 - 54   2008年10月

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    記述言語:英語  

    D-Serine is the endogenous ligand for the glycine binding site of the N-methyl-D-aspartate (NMDA)-type glutamate receptor (GluR) channel and is involved in the regulation of synaptic plasticity, neural network formation, and neurodegenerative disorders. D-Serine is synthesized from L-serine by serine racemase (SR), which was first reported to be localized in astrocytes. However, recently, SR mRNA and its protein have been detected in neurons. In this study, we examined the SR distribution in the brain during postnatal development and in cultured cells by using novel SR knockout mice as negative controls. We found that SR is predominantly localized in pyramidal neurons in the cerebral cortex and hippocampal CA1 region. Double immunofluorescence staining revealed that SR signals colocalized with those of the neuron-specific nuclear protein, but not with the astrocytic markers glial fibrillary acid protein and 3-phosphoglycerate dehydrogenase. In the striatum, we observed SR expression in gamma-aminobutyric acid (GABA)ergic medium-spiny neurons. Furthermore, in the adult cerebellum, we detected weak but significant SR signals in GABAergic Purkinje cells. From these findings, we conclude that SR is expressed predominantly in many types of neuron in the brain and plays a key role in the regulation of brain functions under physiological and pathological conditions via the production of the neuromodulator D-serine.

    DOI: 10.1002/cne.21822

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  • Abundant distribution of TARP gamma-8 in synaptic and extrasynaptic surface of hippocampal neurons and its major role in AMPA receptor expression on spines and dendrites. 査読 国際誌

    Masahiro Fukaya, Mika Tsujita, Maya Yamazaki, Etsuko Kushiya, Manabu Abe, Kaori Akashi, Rie Natsume, Masanobu Kano, Haruyuki Kamiya, Masahiko Watanabe, Kenji Sakimura

    The European journal of neuroscience   24 ( 8 )   2177 - 90   2006年10月

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    記述言語:英語  

    Transmembrane alpha-amino-3-hydroxyl-5-isoxazolepropionate (AMPA) receptor regulatory proteins (TARPs) play pivotal roles in AMPA receptor trafficking and gating. Here we examined cellular and subcellular distribution of TARP gamma-8 in the mouse brain. Immunoblot and immunofluorescence revealed the highest concentration of gamma-8 in the hippocampus. Immunogold electron microscopy demonstrated dense distribution of gamma-8 on the synaptic and extrasynaptic surface of hippocampal neurons with very low intracellular labeling. Of the neuronal surface, gamma-8 was distributed at the highest level on asymmetrical synapses of pyramidal cells and interneurons, whereas their symmetrical synapses selectively lacked immunogold labeling. Then, the role of gamma-8 in AMPA receptor expression was pursued in the hippocampus using mutant mice defective in the gamma-8 gene. In the mutant cornu ammonis (CA)1 region, synaptic and extrasynaptic AMPA receptors on dendrites and spines were severely reduced to 35-37% of control levels, whereas reduction was mild for extrasynaptic receptors on somata (74%) and no significant decrease was seen for intracellular receptors within spines. In the mutant CA3 region, synaptic AMPA receptors were reduced mildly at asymmetrical synapses in the stratum radiatum (67% of control level), and showed no significant decrease at mossy fiber-CA3 synapses. Therefore, gamma-8 is abundantly distributed on hippocampal excitatory synapses and extrasynaptic membranes, and plays an important role in increasing the number of synaptic and extrasynaptic AMPA receptors on dendrites and spines, particularly, in the CA1 region. Variable degrees of reduction further suggest that other TARPs may also mediate this function at different potencies depending on hippocampal subregions, input sources and neuronal compartments.

    DOI: 10.1111/j.1460-9568.2006.05081.x

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  • Distribution of TARP Gamma-8 on hippocampal neurons and its key role in synaptic and extrasynaptic expression for AMPA receptors 査読

    Masahiro Fukaya, Mika Tsujita, Maya Yamazaki, Etsuko Kushiya, Manabu Abe, Kaori Akashi, Masanobu Kano, Haruyuki Kamiya, Kenji Sakimura, Masahiko Watanabe

    NEUROSCIENCE RESEARCH   55   S172 - S172   2006年

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    記述言語:英語   出版者・発行元:ELSEVIER IRELAND LTD  

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  • NMDA receptor GluRepsilon/NR2 subunits are essential for postsynaptic localization and protein stability of GluRzeta1/NR1 subunit. 査読 国際誌

    Manabu Abe, Masahiro Fukaya, Takeshi Yagi, Masayoshi Mishina, Masahiko Watanabe, Kenji Sakimura

    The Journal of neuroscience : the official journal of the Society for Neuroscience   24 ( 33 )   7292 - 304   2004年8月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:SOC NEUROSCIENCE  

    In NMDA receptors, GluRepsilon/NR2 subunits strictly require the GluRzeta1/NR1 subunit to exit from endoplasmic reticulum (ER) to the cell surface in vitro and to the postsynapse in vivo, whereas C terminus-dependent self-surface delivery has been demonstrated for the GluRzeta1 subunit in vitro. To test whether this leads to C terminus-dependent self-postsynaptic expression in neurons in vivo, we investigated the GluRzeta1 subunit in cerebellar granule cells lacking two major GluRepsilon subunits, GluRepsilon1/NR2A and GluRepsilon3/NR2C. In the mutant cerebellum, synaptic labeling for the GluRzeta1 subunit containing the C2 (GluRzeta1-C2) or C2' (GluRzeta1-C2') cassette was reduced at mossy fiber-granule cell synapses to the extrasynaptic level. The loss was not accompanied by decreased transcription and translation levels, increased extrasynaptic labeling, or ER accumulation. Quantitative immunoblot revealed substantial reductions in the mutant cerebellum of GluRzeta1-C2 and GluRzeta1-C2'. The most severe deficit was observed in the postsynaptic density (PSD) fraction: mutant levels relative to the wild-type level were 12.3 +/- 3.3% for GluRzeta1-C2 and 17.0 +/- 4.6% for GluRzeta1-C2'. The GluRzeta1 subunit carrying the C1 cassette (GluRzeta1-C1) was, although low in cerebellar content, also reduced to 12.7 +/- 3.5% in the mutant PSD fraction. Considering a trace amount of other GluRepsilon subunits in the mutant cerebellum, the severe reductions thus represent that the GluRzeta1 subunit, by itself, is virtually unable to accumulate at postsynaptic sites, regardless of C-terminal forms. By protein turnover analysis, the degradation of the GluRzeta1 subunit was accelerated in the mutant cerebellum, being particularly rapid for that carrying the C2 cassette. Therefore, accompanying expression of GluRepsilon subunits is essential for postsynaptic localization and protein stability of the GluRzeta1 subunit.

    DOI: 10.1523/JNEUROSCI.1261-04.2004

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  • Differential palmitoylation of two mouse glutamate receptor interacting protein 1 forms with different N-terminal sequences 査読

    M Yamazaki, M Fukaya, M Abe, K Ikeno, T Kakizaki, M Watanabe, K Sakimura

    NEUROSCIENCE LETTERS   304 ( 1-2 )   81 - 84   2001年5月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ELSEVIER SCI IRELAND LTD  

    Glutamate receptor interacting protein (GRIP) is a member of the PDZ domain-containing protein family that is localized in the postsynaptic density area. This protein has been reported to interact specifically with the C-termini of AMPA-selective glutamate receptor channel subunits, GluR alpha2 and GluR alpha3 through its PDZ domains. To clarify the physiological functions of GRIP, we cloned mouse GRIP1, and found that there are three sites for alternative splicing and two putative translational start codons by characterizing GRIP1 cDNA clones and reverse transcription-polymerase chain reaction products. Metabolic labeling of COS-7 cells expressing two N-terminal GRIP1 proteins demonstrated that these proteins differed in their pattern of palmitoylation. These findings suggested that the molecular diversity of GRIP1 underlies the localization and functional heterogeneity of this protein. (C) 2001 Elsevier Science Ireland Ltd. All rights reserved.

    DOI: 10.1016/S0304-3940(01)01766-9

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  • Purkinje cell-specific and inducible gene recombination system generated from C57BL/6 mouse ES cells 査読

    K Kitayama, M Abe, T Kakizaki, D Honma, R Natsume, M Fukaya, M Watanabe, J Miyazaki, M Mishina, K Sakimura

    BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS   281 ( 5 )   1134 - 1140   2001年3月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ACADEMIC PRESS INC  

    Spatiotemporally restricted gene targeting is needed for analyzing the functions of various molecules in a variety of biological phenomena. We have generated an inducible cerebellar Purkinje cell-specific gene targeting system. This was achieved by establishing a mutant mouse line (D2CPR) from a C57BL/6 mouse ES cell line, which expressed a fusion protein consisting of the Cre recombinase and the progesterone receptor (CrePR). The Purkinje cell-specific expression of CrePR was attained by inserting CrePR into the glutamate receptor delta2 subunit (GluR delta2) gene, which was expressed specifically in the Purkinje cells. Using the transgenic mice carrying the Cre-mediated reporter gene, we showed that the antiprogesterone RU486 could induce recombinase activity of the CrePR protein specifically in the mature cerebellar Purkinje cells of the D2CPR line. Thus this mutant line will be a useful tool for studying the molecular function of mature Purkinje cells by manipulating gene expression in a temporally restricted manner. (C) 2001 Academic Press.

    DOI: 10.1006/bbrc.2001.4492

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  • Absence of prostaglandin E-2-induced hyperalgesia in NMDA receptor epsilon subunit knockout mice 査読

    T Minami, J Sugatani, K Sakimura, M Abe, M Mishina, S Ito

    BRITISH JOURNAL OF PHARMACOLOGY   120 ( 8 )   1522 - 1526   1997年4月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:STOCKTON PRESS  

    1 We have previously found that intrathecal administration of prostaglandins E-2 (PGE(2)) and D-2 (PGD(2)) into conscious mice induced hyperalgesia by the hut plate test, The present study investigated the involvement of N-methyl-D-aspartate (NMDA) receptor in the prostaglandin-induced hyperalgesia by use of mice lacking NMDA receptor epsilon 1, epsilon 4, Or epsilon 1/epsilon 4 subunits,
    2 PGE(2) induced hyperalgesia over a wide rang of doses from 50 pg to 500 ng kg (1)in wild-type mice, But PGE(2) could not induce hyperalgesia. in epsilon 1, epsilon 4, Or epsilon 1/epsilon 4 subunit knockout mice;
    3 The NMDA receptor antagonist D-(-)-2-amino-5-phosphonovaleric acid (D-AP5), the nan-NP IDA receptor antagonist gamma-D-glutamylaminomethyl sulphonic acid (GAMS), and the nitric oxide synthase inhibitor N-omega-nitro-L-arginine methyl ester (L-NAME) inhibited the PGE(2)-induced hyperalgesia in wild-type mice.
    4 PGD(2) induced hyperalgesia al doses of 25 ng to 250 ng kg(-1) in both wild-type and epsilon 1/epsilon 4 subunit knockout mice. The substance P receptor antagonist CP 96,345 blocked the PGD(2)-induced hyperalgesia in wild-type and epsilon 1/epsilon 4 subunit knockout mice.
    5 These results demonstrate that the pathways leading to hyperalgesia are different; between PGD(2) and PGE(2).and that both epsilon 1 and epsilon 4 subunits of the NMDA receptor are involved in the PGE(2)-induced hyperalgesia.

    DOI: 10.1038/sj.bjp.0701067

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MISC

  • 胚盤胞補完法とES細胞を用いた甲状腺再生

    冉慶松, 周啓亮, 小田加奈子, 泰江彰浩, 阿部学, 笹岡俊邦, 崎村健司, 味岡洋一, 西條康夫

    日本再生医療学会総会(Web)   20th   2021年

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  • 胚盤胞補完法とES細胞を用いた甲状腺再生

    冉慶松, 周けい亮, 叶許緑, 小田加奈子, 泰江章博, 阿部学, 崎村建司, 笹岡俊邦, 味岡洋一, 西條康夫

    日本再生医療学会総会(Web)   19th   2020年

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  • CRISPR遺伝子編集系を電気穿孔法で受精卵に導入して作製したNanos3ノックアウトマウス生殖巣の形態解析 査読

    大平 拓也, 中務 胞, 阿部 学, 夏目 理恵, 杉村 智史, 崎村 建司, 山城 秀昭

    日本繁殖生物学会 講演要旨集   112 ( 0 )   P - 106-P-106   2019年

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    記述言語:日本語   出版者・発行元:日本繁殖生物学会  

    <p>【目的】本研究は,生殖細胞の形成に不可欠であるNanos3をノックアウトしたマウスの生殖巣を解析することを目的に,CRISPR/Cas9システムを電気穿孔法で受精卵に導入するTAKE法とGONAD法での変異導入効率の違いを胚盤胞で調べた。さらに遺伝子編集を惹起した受精卵より産仔を得て,生殖巣の解析を行った。【方法】TAKE法:4週齢以上のマウスに過排卵処理を施して得た前核期卵子は,Cas9mRNAとgRNAを満たしたチャンバー内に並べ,電気穿孔を行った。GONAD法:10週齢以上の交配させたマウスの卵管を体外に出し,Cas9とgRNAを卵管内に注入し電極を用いて電気穿孔法を施した。TAKE法およびGONAD法にて得られた産仔の尾または胚盤胞からDNAを抽出し,PCRおよびsquenceにてhomo,hetero,wildに分類した。産仔は10週齢で生殖巣を採取し,生殖巣表現型の差異を比較した。生殖巣は,H&E染色により組織形態学的解析を行った。【結果】TAKE法の胚盤胞区では,17.6%(homo:2個/ 5.9%, hetero:4個/11.7%),移植区では18.5%(homo:6匹/ 17.2%, hetero:1匹/ 1.3%)の効率で変異を確認できた。GONAD法では,18.6%の効率で変異が確認でき,全てhomo欠損個体であった。精巣および卵巣は,wild個体とhomo欠損個体でサイズに著しく差が生じていた。H&E染色の結果,homo欠損個体では,精子を含む生殖細胞は確認されず,セルトリ細胞のみが確認された。卵巣のH&E染色像を観察した結果,homo欠損個体において卵子および卵胞は確認されなかった。以上,電気穿孔法で受精卵での遺伝子編集により,TAKE法とGONAD法では同程度の効率でNanos3遺伝子への変異を導入することが可能であり,かつ作製したNanos3 KOマウスの生殖巣は,精巣の精細管内には精細胞が形成されず,卵巣内にも卵胞,卵母細胞が形成されないことを明らかにした。</p>

    DOI: 10.14882/jrds.112.0_P-106

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  • 胚盤胞補完法とES細胞を用いたマウス生体内における肺臓器再生

    冉慶松, 周啓亮, 北原哲彦, 叶許緑, 佐々木健太, 齋木琢郎, 松本吉史, 森山雅人, 泰江章博, 阿部学, 味岡洋一, 笹岡俊邦, 西條康夫

    日本再生医療学会総会(Web)   18th   2019年

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  • CRF‐Venus Δ Neoマウスの分界条床核外側部CRHニューロンにおけるシナプス入力の性差の電気生理学的な検討―ホルマリンテストを用いて

    萩原裕子, 崎村建司, 阿部学, 井樋慶一, 明間立雄, 舩橋利也

    日本神経内分泌学会学術集会プログラム・抄録集   45th   43   2018年

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    記述言語:日本語  

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  • RNA顆粒構成因子RNG105は、シナプス後部形態の可塑的変化を制御し、記憶形成に必要である

    中山 啓, 阿部 学, 山崎 真弥, 藤川 顕寛, 野田 昌晴, 二木 啓, 御子柴 克彦, 崎村 建司, 椎名 伸之

    生命科学系学会合同年次大会   2017年度   [1P - 1301]   2017年12月

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    記述言語:英語   出版者・発行元:生命科学系学会合同年次大会運営事務局  

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  • GLUD1 deficient mouse as a model animal of depressionlike behavior

    K. Sakimura, C. Nakamoto, M. Abe, M. Kawamura, H. Uchida, M. Watanabe, M. Kano

    JOURNAL OF NEUROCHEMISTRY   142   203 - 203   2017年8月

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    記述言語:英語   掲載種別:研究発表ペーパー・要旨(国際会議)   出版者・発行元:WILEY  

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  • 副腎不全に伴う視床下部室傍核CRFニューロンにおけるバゾプレシン発現について

    山形聡, TALUKDER Ashraf Hossain, 内田克哉, 佐藤達也, 夏目里恵, 阿部学, 崎村建司, 井樋慶一

    バゾプレシン研究会プログラム・講演抄録   27th   23   2016年12月

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    記述言語:日本語  

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  • 視床下部コルチコトルピン放出因子(CRF)ニューロンへの抑制性入力とセロトニン(5‐HT)による制御機序に関する研究

    佐藤隆幸, 菅谷琢磨, 福田敦夫, 杉本直哉, 布施俊光, 内田克哉, 阿部学, 山崎真弥, 崎村建司, 井樋慶一, 井樋慶一

    日本内分泌学会雑誌   91 ( 2 )   526   2015年9月

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    記述言語:日本語  

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  • A unique western blot method to measure the number of glutamate receptor subunits

    K. Sakimura, C. Nishikawa, I. Watanabe, C. Nakamoto, Y. Suzuki, H. Azechi, R. Natsume, M. Abe, M. Watanabe, M. Kano

    JOURNAL OF NEUROCHEMISTRY   134   163 - 163   2015年8月

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    記述言語:英語   掲載種別:研究発表ペーパー・要旨(国際会議)   出版者・発行元:WILEY-BLACKWELL  

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  • Proteasome Dysfunction Induces Muscle Growth Defects And Protein Aggregation

    Yasuo Kitajima, Naoki Suzuki, Yoshitaka Tashiro, Hitoshi Warita, Masaaki Kato, Maki Tateyama, Risa Ando, Rumiko Izumi, Maya Yamazaki, Manabu Abe, Kenji Sakimura, Hidefumi Ito, Urushitani Makoto, Ryoichi Nagatomi, Ryosuke Takahashi, Masashi Aoki

    MEDICINE AND SCIENCE IN SPORTS AND EXERCISE   46 ( 5 )   352 - 352   2014年5月

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    記述言語:英語   掲載種別:研究発表ペーパー・要旨(国際会議)   出版者・発行元:LIPPINCOTT WILLIAMS & WILKINS  

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  • 骨格筋特異的プロテアソーム機能不全は筋成長不全ならびに異常タンパク質の蓄積を引き起こす

    北嶋康雄, 鈴木直輝, 田代善崇, 割田仁, 加藤昌昭, 竪山真規, 安藤里紗, 井泉瑠美子, 山崎真弥, 阿部学, 崎村建司, 伊東秀文, 漆谷真, 永富良一, 高橋良輔, 青木正志

    体力科学   63 ( 1 )   142   2014年2月

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    記述言語:日本語  

    DOI: 10.7600/jspfsm.63.142

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  • 視索前野のGABA作動性神経とオレキシン神経の機能的結合について(16.神経回路・脳の情報処理,ポスター,日本生物物理学会年会第51回(2013年度))

    Kanda(Tsujino),Natsuko, Saito,Yuki, Abe,Manabu, Sakimura,Kenji, Yanagisawa,Masashi, Sakurai,Takeshi

    生物物理   53 ( 1 )   S197   2013年9月

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    記述言語:日本語   出版者・発行元:一般社団法人日本生物物理学会  

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  • Nna1 is related to motor learning and dynamics of glutamate receptor subunits

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    記述言語:英語   掲載種別:研究発表ペーパー・要旨(国際会議)   出版者・発行元:WILEY-BLACKWELL  

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  • Branching Kinase (BrancK) regulates arborization of neurites.

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    記述言語:英語   掲載種別:研究発表ペーパー・要旨(国際会議)   出版者・発行元:AMER SOC CELL BIOLOGY  

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  • 脊髄運動神経特異的26Sプロテアソームサブユニット欠損マウスはALS的病理所見を伴う運動神経細胞死を呈する

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  • 神経変性疾患に関する調査研究 マウス26Sプロテアソーム欠損を用いた神経変性疾患モデルの検討

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  • 骨格筋特異的プロテアソーム機能不全は筋萎縮ならびに異常タンパク質の蓄積を引き起こす

    北嶋康雄, 鈴木直輝, 田代善崇, 割田仁, 加藤昌昭, 竪山真規, 安藤里紗, 井泉瑠美子, 山崎真弥, 阿部学, 崎村建司, 伊東秀文, 漆谷真, 永富良一, 高橋良輔, 青木正志

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  • 神経変性疾患モデル作製のための26Sプロテアソームコンディショナルノックアウトマウスの確立と解析

    田代 善崇, 伊東 秀文, 井上 治久, 山崎 真弥, 阿部 学, 三澤 日出巳, 崎村 建司, 高橋 良輔

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  • Differential regulation between duplicated genes in protocadherin-alpha cluster acquired without subsequent mutations

    Ryosuke Kaneko, Manabu Abe, Takahiro Hirabayashi, Arikuni Uchimura, Kenji Sakimura, Yuchio Yanagawa, Takeshi Yagi

    NEUROSCIENCE RESEARCH   71   E46 - E46   2011年

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    記述言語:英語   掲載種別:研究発表ペーパー・要旨(国際会議)   出版者・発行元:ELSEVIER IRELAND LTD  

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  • The analysis of 26S proteasome conditional knockout mice for the mechanisms of neurodegenerative diseases

    Yoshitaka Tashiro, Hidehumi Ito, Haruhisa Inoue, Maya Yamazaki, Manabu Abe, Hidemi Misawa, Kenji Sakimura, Ryosuke Takahashi

    NEUROSCIENCE RESEARCH   71   E392 - E392   2011年

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    記述言語:英語   掲載種別:研究発表ペーパー・要旨(国際会議)   出版者・発行元:ELSEVIER IRELAND LTD  

    DOI: 10.1016/j.neures.2011.07.1719

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  • 神経変性疾患に関する調査研究 マウス26Sプロテアソーム欠損を用いた神経変性疾患モデルの検討

    高橋良輔, 高橋良輔, 田代善崇, 田代善崇, 伊東秀文, 井上治久, 井上治久, 山崎真弥, 阿部学, 三澤日出巳, 崎村建司

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  • Glutamate receptor delta 2 is essential for input pathway-dependent regulation of synaptic AMPAR contents in cerebellar Purkinje cells 査読

    Yamasaki Miwako, Miyazaki Taisuke, Azechi Hirotsugu, Abe Manabu, Natsume Rie, Hagiwara Teruki, Aiba Atsu, Mishina Masayoshi, Sakimura Kenji, Watanabe Masahiko

    NEUROSCIENCE RESEARCH   71   E93   2011年

  • Quantitative analysis of kainate receptor subunits in the mouse brain 査読

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    NEUROSCIENCE RESEARCH   71   E315 - E316   2011年

  • Synaptic localization and content of four AMPA receptor subunits at excitatory hippocampal synapses 査読

    Miwako Yamasaki, Masahiro Fukaya, Manabu Abe, Kenji Sakimura, Masahiko Watanabe

    NEUROSCIENCE RESEARCH   68   E56 - E56   2010年

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    記述言語:英語   掲載種別:研究発表ペーパー・要旨(国際会議)   出版者・発行元:ELSEVIER IRELAND LTD  

    DOI: 10.1016/j.neures.2010.07.015

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  • Analysis of physiological functions of cerebellar AMPA receptor subunits using a Purkinje cell-selective gene targeting system

    Manabu Abe, Kouichi Hashimoto, Taisuke Miyazaki, Hirotsugu Azechi, Rie Natsume, Masahiko Watanabe, Masanobu Kano, Kenji Sakimura

    NEUROSCIENCE RESEARCH   68   E231 - E231   2010年

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    記述言語:英語   掲載種別:研究発表ペーパー・要旨(国際会議)   出版者・発行元:ELSEVIER IRELAND LTD  

    DOI: 10.1016/j.neures.2010.07.1019

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  • Behavioral characteristics of CLICK-III/CaMKIgamma knockout male mice in social and non-social context

    M. Nakata, S. Takemoto-Kimura, N. Akashi, T. Sakamoto, M. Kakeyama, M. Yamazaki, M. Abe, R. Natsume, K. Sakimura, T. Takeuchi, H. Mori, M. Mishina, H. Bito, S. Ogawa

    NEUROSCIENCE RESEARCH   68   E413 - E414   2010年

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    記述言語:英語   掲載種別:研究発表ペーパー・要旨(国際会議)   出版者・発行元:ELSEVIER IRELAND LTD  

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  • The establishment and analysis of 26S proteasome conditional knockout mice for the mechanisms of neurodegenerative diseases

    Yoshitaka Tashiro, Haruhisa Inoue, Maya Yamazaki, Manabu Abe, Hidehumi Ito, Hidemi Misawa, Kenji Sakimura, Ryosuke Takahashi

    NEUROSCIENCE RESEARCH   68   E96 - E96   2010年

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    記述言語:英語   掲載種別:研究発表ペーパー・要旨(国際会議)   出版者・発行元:ELSEVIER IRELAND LTD  

    DOI: 10.1016/j.neures.2010.07.187

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  • Gene duplication alters expression and DNA methylation in Protocadherin-alpha cluster

    Ryosuke Kaneko, Manabu Abe, Takahiro Hirabayashi, Arikuni Uchimura, Kenji Sakimura, Yuchio Yanagawa, Takeshi Yagi

    NEUROSCIENCE RESEARCH   68   E352 - E353   2010年

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    記述言語:英語   掲載種別:研究発表ペーパー・要旨(国際会議)   出版者・発行元:ELSEVIER IRELAND LTD  

    DOI: 10.1016/j.neures.2010.07.1562

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  • Histological and behavioral analyses in CL3/CaMKIgamma-deficient mice

    Sayaka Takemoto-Kimura, Satoshi Kamijo, Shinichiro Horigane, Kanzo Suzuki, Hiroyuki Okuno, Asahi Haijima, Toshihiro Endo, Tomonori Takeuchi, Manabu Abe, Rie Natsume, Maya Yamazaki, Hisashi Mori, Chiharu Tohyama, Kenji Sakimura, Masayoshi Mishina, Masaki Kakeyama, Haruhiko Bito

    NEUROSCIENCE RESEARCH   68   E404 - E405   2010年

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    記述言語:英語   掲載種別:研究発表ペーパー・要旨(国際会議)   出版者・発行元:ELSEVIER IRELAND LTD  

    DOI: 10.1016/j.neures.2010.07.1795

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  • 2-Arachidonoylglycerol produced by diacylglycerol lipase alpha mediates retrograde suppression of synaptic transmission in the CNS

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    NEUROSCIENCE RESEARCH   68   E88 - E88   2010年

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    記述言語:英語   掲載種別:研究発表ペーパー・要旨(国際会議)   出版者・発行元:ELSEVIER IRELAND LTD  

    DOI: 10.1016/j.neures.2010.07.153

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  • 26Sプロテアソーム部位特異的欠損マウスを用いた神経変性疾患モデルの確立

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  • 神経変性疾患に関する調査研究 マウス26Sプロテアソーム欠損による神経変性疾患モデルの確立と解析

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  • NMDA RECEPTOR GLUN2B SUBUNIT IS CRUCIAL FOR CHANNEL FUNCTION AND POST-SYNAPTIC MOLECULAR ORGANIZATION AT HIPPOCAMPAL CA3 SYNAPSES

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  • 神経変性疾患に関する調査研究 マウス26Sプロテアソーム欠損による神経変性疾患モデルの確立

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  • Loss of cerebellar granule cell AMPA receptors contributes to ataxia in stargazer mice

    May Yamazaki, Masahiro Fukaya, Taisuke Miyazaki, Manabu Abe, Rie Natsume, Masahiko Watanabe, Kenji Sakimura

    NEUROSCIENCE RESEARCH   65   S140 - S140   2009年

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    記述言語:英語   掲載種別:研究発表ペーパー・要旨(国際会議)   出版者・発行元:ELSEVIER IRELAND LTD  

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  • Roles of clustered genomic organization of Protocadherin-alpha on individual neuron specific Protocadherin-alpha choice

    Ryosuke Kaneko, Manabu Abe, Takahiro Hirabayashi, Arikuni Uchimura, Kenji Sakimura, Yuchio Yanagawa, Takeshi Yagi

    NEUROSCIENCE RESEARCH   65   S89 - S89   2009年

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    記述言語:英語   掲載種別:研究発表ペーパー・要旨(国際会議)   出版者・発行元:ELSEVIER IRELAND LTD  

    DOI: 10.1016/j.neures.2009.09.365

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  • The establishment and analysis of 26S proteasome conditional knockout mice for the mechanisms of neurodegenerative diseases

    Yoshitaka Tashiro, Haruhisa Inoue, Maya Yamazaki, Manabu Abe, Hidemi Misawa, Kenji Sakimura, Ryosuke Takahashi

    NEUROSCIENCE RESEARCH   65   S248 - S248   2009年

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    記述言語:英語   掲載種別:研究発表ペーパー・要旨(国際会議)   出版者・発行元:ELSEVIER IRELAND LTD  

    DOI: 10.1016/j.neures.2009.09.1407

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  • GluR epsilon 2 subunit is crucial for NMDA receptor activity and regulation of postsynaptic macromolecular organization 査読

    Kaori Akashi, Toshikazu Kakizaki, Haruyuki Kamiya, Masahiro Fukaya, Miwako Yamasaki, Manabu Abe, Masahiko Watanabe, Kenji Sakimura

    NEUROSCIENCE RESEARCH   65   S139 - S140   2009年

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    記述言語:英語   掲載種別:研究発表ペーパー・要旨(国際会議)   出版者・発行元:ELSEVIER IRELAND LTD  

    DOI: 10.1016/j.neures.2009.09.693

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  • [New generation technique of gene targeting: its problem and prospect].

    Kenji Sakimura, Manabu Abe, Maya Yamazaki, Hiroshi Usui

    Tanpakushitsu kakusan koso. Protein, nucleic acid, enzyme   53 ( 11 )   1383 - 91   2008年9月

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    記述言語:日本語  

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  • 神経変性疾患モデル作製のための26Sプロテアソームコンディショナルノックアウトマウスの確立と解析

    田代善崇, 井上治久, 山崎真弥, 阿部学, 村松一洋, 原田彰宏, 三澤日出巳, 崎村建司, 高橋良輔, 高橋良輔

    生化学   3P-1105   2008年

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  • The establishment of 26S proteasome conditional knockout mice for exploring the mechanisms of neurodegenerative diseases

    Yoshitaka Tashiro, Haruhisa Inoue, Maya Yamazaki, Manabu Abe, Hidemi Misawa, Kenji Sakimura, Ryosuke Takahashi

    NEUROSCIENCE RESEARCH   61   S124 - S124   2008年

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    記述言語:英語   掲載種別:研究発表ペーパー・要旨(国際会議)   出版者・発行元:ELSEVIER IRELAND LTD  

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  • Transmembrane AMPA receptor regulatory protein gamma-8 is involved with the regulation of spontaneous activity and mental condition

    Maya Yamazaki, Masahiro Fukaya, Manabu Abe, Rie Natsume, Masahiko Watanabe, Kenji Sakimura

    NEUROSCIENCE RESEARCH   61   S208 - S208   2008年

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    記述言語:英語   掲載種別:研究発表ペーパー・要旨(国際会議)   出版者・発行元:ELSEVIER IRELAND LTD  

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  • 神経部位特異的26Sプロテアソームサブユニット欠損マウスの作製

    田代 善崇, 山崎 真弥, 阿部 学, 崎村 建司, 小松 雅明, 田中 啓二, 高橋 良輔

    臨床神経学   47 ( 12 )   1084 - 1084   2007年12月

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    記述言語:日本語   出版者・発行元:(一社)日本神経学会  

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  • 新世代のジーンターゲティング : その問題点と展望

    崎村建司, 阿部学, 山崎真弥, 薄井宏

    生化学   79 ( 4 )   340 - 349   2007年4月

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    出版者・発行元:日本生化学会  

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  • [New-generation gene targeting technique: its problems and prospects].

    Kenji Sakimura, Manabu Abe, Maya Yamazaki, Hiroshi Usui

    Seikagaku. The Journal of Japanese Biochemical Society   79 ( 4 )   340 - 9   2007年4月

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    記述言語:日本語  

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  • Systematic generation of conditional knockout mice using C57BL/6 ES cell RENKA

    Kenji Sakimura, Manabu Abe, Maya Yamazaki, Hiroshi Usui

    NEUROSCIENCE RESEARCH   58   S30 - S30   2007年

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    記述言語:英語   掲載種別:研究発表ペーパー・要旨(国際会議)   出版者・発行元:ELSEVIER IRELAND LTD  

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  • Effects of GluR beta 2/GluR6 disruption on the dynamics of other kainate receptor subunits

    Kaori Akashi, Manabu Abe, Toshikazu Kakizaki, Rie Natsume, Kenji Sakimura

    NEUROSCIENCE RESEARCH   55   S82 - S82   2006年

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    記述言語:英語   掲載種別:研究発表ペーパー・要旨(国際会議)   出版者・発行元:ELSEVIER IRELAND LTD  

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  • Stargazin family gamma 4 is involved in the dynamics of AMPA receptor expression

    Miharu Komai, Maya Yamazaki, Mika Tsujita, Manabu Abe, Rie Natsume, Kenji Sakimura

    NEUROSCIENCE RESEARCH   55   S139 - S139   2006年

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    記述言語:英語   掲載種別:研究発表ペーパー・要旨(国際会議)   出版者・発行元:ELSEVIER IRELAND LTD  

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  • Generation of TARP gamma-2 (stargazin) conditional knockout mouse

    PH Lee, M Abe, M Yamazaki, M Tsujita, R Natsume, K Sakimura

    JOURNAL OF PHARMACOLOGICAL SCIENCES   100   246P - 246P   2006年

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    記述言語:英語   掲載種別:研究発表ペーパー・要旨(国際会議)   出版者・発行元:JAPANESE PHARMACOLOGICAL SOC  

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  • The role of TARP gamma-8 (DVCC gamma-8) in AMPA receptor localization and function in hippocampus

    M Tsujita, M Fukaya, H Kamiya, M Abe, M Yamazaki, E Kushiya, R Natsume, M Watanabe, K Sakimura

    JOURNAL OF PHARMACOLOGICAL SCIENCES   100   76P - 76P   2006年

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    記述言語:英語   掲載種別:研究発表ペーパー・要旨(国際会議)   出版者・発行元:JAPANESE PHARMACOLOGICAL SOC  

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  • Generation and biochemical analysis of a GluR alpha 2 knockout mouse

    Hirotsugu Azechi, Manabu Abe, Rie Natsume, Kenji Sakimura

    NEUROSCIENCE RESEARCH   55   S80 - S80   2006年

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    記述言語:英語   掲載種別:研究発表ペーパー・要旨(国際会議)   出版者・発行元:ELSEVIER IRELAND LTD  

    Web of Science

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  • Purkinje cell-specific gene recombination mouse line generated from C57BL/6 strain ES cells

    T Hagiwara, M Tsujita, M Abe, K Akashi, R Natsume, K Sakimura

    JOURNAL OF PHARMACOLOGICAL SCIENCES   97   283P - 283P   2005年

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    記述言語:英語   掲載種別:研究発表ペーパー・要旨(国際会議)   出版者・発行元:JAPANESE PHARMACOLOGICAL SOC  

    Web of Science

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  • 興奮毒性におけるNMDA型受容体の関与

    夏目里恵, 阿部学, 菅井智昭, 葉山文恵, 崎村建司

    新潟医学会雑誌   119 ( 12 )   730 - 734   2005年

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    出版者・発行元:新潟大学  

    グルタミン酸興奮毒性は,脳虚血やてんかんなどの急性疾患ばかりでなく,長い過程を経る神経変性疾患による神経細胞死の原因としても注目されている.グルタミン酸受容体チャネルは,興奮毒性発現機序において中心的な役割を果たしていると考えられてきた.とりわけ,高いCaイオンの透過性を持つNMDA型グルタミン酸受容体はその鍵を握る分子として注目されてきた.この受容体の機能特性を決定する4種類のGluRεサブユニットノックアウトマウスを用いて,カイニン酸急性毒性におけるNMDA型受容体の関与を検証した.その結果,GluRε1サブユニットがカイニン酸による興奮毒性発現に最も重要な役割を果たしていることが明らかになった.さらに,小脳顆粒細胞に有意に発現するGluRε3や幼者期に主な発現があるGluRε4にも毒性発現への影響力が有ることから,NMDA型受容体は様々な機序で興奮毒性に関与していることが示唆された.

    CiNii Article

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共同研究・競争的資金等の研究

  • 加齢に伴う老化細胞蓄積メカニズムとその病的老化形質に対する関与の解明

    2019年10月 - 2025年3月

    国立研究開発法人日本医療研究開発機構  革新的先端研究開発支援事業 AMED-CREST「全ライフコースを対象とした個体の機能低下機構の解明」 

    南野徹, 奥田修二郎

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    担当区分:研究分担者 

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  • 細胞内シグナル伝達系の光操作による革新的シナプス可塑性介入技術の研究開発

    2019年4月 - 2024年3月

    国立研究開発法人日本医療研究開発機構  日本医療研究開発機構研究費 革新的技術による脳機能ネットワークの全容解明プロジェクト」技術開発個別課題(チーム型) 

    濱田駿, 渡部文子

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    担当区分:研究分担者 

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  • コンディショナル変異ラットを用いた記憶保持の強化の分子機構解明

    研究課題/領域番号:18KK0458  2019年4月 - 2021年3月

    日本学術振興会  科学研究費助成事業 国際共同研究加速基金(国際共同研究強化(A))  国際共同研究加速基金(国際共同研究強化(A))

    阿部 学

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    担当区分:研究代表者 

    配分額:15600000円 ( 直接経費:12000000円 、 間接経費:3600000円 )

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  • 一世代コンディショナル変異導入法を用いた神経疾患原因遺伝子の生理機能解析

    研究課題/領域番号:17K01972  2017年4月 - 2021年3月

    日本学術振興会  科学研究費助成事業 基盤研究(C)  基盤研究(C)

    阿部 学

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    配分額:4680000円 ( 直接経費:3600000円 、 間接経費:1080000円 )

    生体内における遺伝子の生理機能解析法の中でも特に強力な技術として、Cre-loxP組換え系を用いたコンディショナルノックアウト法が発展してきた。その原理を利用した遺伝子発現系として、ノックイン-トランスジェニック型でCre組換え依存的に強力なプロモーターにより遺伝子を発現させるシステムや、数種のloxP 類縁配列と組み合わせDNA配列を逆位にすることで遺伝子発現を制御するFLEx(またはDIO)と呼ばれるシステムなども存在するが、多くの場合は内在の遺伝子構造と異なっていることが原因で、正確な生理機能評価が難しいことが問題となっている。研究代表者は、FLExシステムと異動物種由来ゲノム等を用いた新規コンディショナル遺伝子発現制御法を開発することにより、この問題を解決できる可能性を見出した。 本研究の目的は、開発された新規遺伝子発現制御法の有効性を確認すると同時に、遺伝子変異-中枢神経系回路発達-表現型の関連性を捉えやすい小脳発達期をモデルとして選択し、生理機能が明らかでない2つの神経疾患原因遺伝子、炭酸脱水素酵素関連タンパクCA8(マウス遺伝子名Car8)および翻訳伸長因子EEF1A2 (Eef1a2)とそれらの関連分子を主な変異導入の対象として、小脳発達の分子機序の一端を明らかにすることである。 一方、従来の遺伝子改変マウス作製が遺伝学、発生工学を中心的技術とするため長い実験期間を要するという最大の問題については、近年著しく発展しているゲノム編集技術を適用することで解決可能だと考えられた。研究代表者は簡便なノックインマウス作製法であるマウス初期胚に対する電気穿孔法を用いることにより、既に一世代-低分子タグノックインマウス作製技術を確立しており、より簡便に長い配列のノックインを可能とするための技術開発も行う。

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  • 乳癌免疫微小環境における脂質メディエーター分子の役割解明と治療応用を目指した研究

    研究課題/領域番号:19H03714  2019年4月 - 2022年3月

    日本学術振興会  科学研究費助成事業 基盤研究(B)  基盤研究(B)

    永橋 昌幸, 奥田 修二郎, 阿部 学, 諸 和樹, 崎村 建司, 五十嵐 道弘, 若井 俊文, 土田 純子

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    配分額:17550000円 ( 直接経費:13500000円 、 間接経費:4050000円 )

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  • S1Pの癌代謝制御機構に着眼したトリプルネガティブ乳癌の病態と薬剤耐性機序の解明

    研究課題/領域番号:18K19576  2018年6月 - 2021年3月

    日本学術振興会  科学研究費助成事業 挑戦的研究(萌芽)  挑戦的研究(萌芽)

    永橋 昌幸, 若井 俊文, 五十嵐 道弘, 崎村 建司, 阿部 学, 奥田 修二郎, 土田 純子

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    配分額:6240000円 ( 直接経費:4800000円 、 間接経費:1440000円 )

    トリプルネガティブ乳癌(TNBC)は生物学的に悪性度が高く、薬物療法への抵抗性を生じやすく予後不良である。近年の網羅的遺伝子解析にも関わらず、TNBCは発癌や癌の悪化に寄与するドライバー遺伝子変異に乏しく、その病態メカニズムは未だ不明の点が多い。スフィンゴシン-1-リン酸(S1P)は、脂質でありながらタンパク質と同じように情報伝達物質として働く脂質メディエーターであり、癌で重要な役割を担っている。私達は、「TNBCの高悪性度の病態は、遺伝子やタンパク質の異常だけでは説明がつかず、その背景には脂質メディエーターを介した癌代謝制御機構が関与している」と仮説を立て、本研究を企画した。本研究の目的は「脂質メディエーター・S1Pを介した癌代謝制御機構に着目してTNBCの病態メカニズムを解明し、新規治療法開発への研究基盤を確立すること」である。本研究実績として、TNBCマウスモデルを用いて、癌の発育進展においてS1Pが重要な役割を果たしていることが分かった。また、臨床検体のリピドミクス解析により、TNBC患者におけるスフィンゴリン脂質の動態について明らかにした。さらにS1Pにより制御される癌の代謝状態について、メタボローム解析を行い、データを収集した。さらに臨床病理学的因子との比較により、TNBCにおけるS1Pの臨床的意義を見出し、今後の治療開発へ向けた研究の礎となることが期待される。

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  • 生体内動態解析を可能とする網羅的タンパク質分子標識法の開発

    研究課題/領域番号:17K19444  2017年6月 - 2019年3月

    日本学術振興会  科学研究費助成事業 挑戦的研究(萌芽)  挑戦的研究(萌芽)

    崎村 建司, 阿部 学

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    配分額:6500000円 ( 直接経費:5000000円 、 間接経費:1500000円 )

    本研究の目的は、生体内で機能するタンパク質分子を正確に検出する方法を新たに開発し、その手法を神経系で機能する分子群に適用することである。このために、抗体に代わる低分子量のケミカルプローブとそれを認識するペプチドタグを組み合わせた系を採用した。その結果、遺伝子編集方法を用いて、ペプチドタグを標的分子の機能に影響しないと予想される部位に迅速に挿入する方法の開発に成功した。この手法を用いて、NMDA型グルタミン酸受容体のサブユニット構造を電子顕微鏡で可視化する金粒子ラベルしたプローブが結合するhD2タグをGluN1分子にノックインしたマウスを樹立した。

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  • 多様な発想で研究可能なリソースとしての遺伝子改変マーモセット作製法の開発

    研究課題/領域番号:16H04650  2016年4月 - 2019年3月

    日本学術振興会  科学研究費助成事業 基盤研究(B)  基盤研究(B)

    崎村 建司, 阿部 学, 夏目 里恵, 中務 胞

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    配分額:16640000円 ( 直接経費:12800000円 、 間接経費:3840000円 )

    ヒト脳機能と疾患を理解する上で必要な霊長類マーモセットの遺伝子改変モデル作製には、経済的・倫理的障壁があり、これまで実現困難であった。本研究では、これら問題を克服するために、安価且つ動物福祉的にもかなった手法でマーモセット受精卵を取得する方法を検討した。廃用個体や死亡個体から取得した卵巣組織をヌードマウスの腎臓被膜下に移植し、ホルモン投与により移植片からの卵子の取得に成功した。さらに、取得した卵子から、桑実胚まで発生する受精卵を作製する方法を見いだした。

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  • 食道癌のリンパ行性進展に関与する脂質メディエーター分子機構の解明および臨床的意義

    研究課題/領域番号:16K10491  2016年4月 - 2019年3月

    日本学術振興会  科学研究費助成事業 基盤研究(C)  基盤研究(C)

    市川 寛, 永橋 昌幸, 若井 俊文, 羽入 隆晃, 小杉 伸一, 宗岡 悠介, 中野 雅人, 田中 花菜, 須藤 翔, 崎村 健司, 阿部 学

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    配分額:4680000円 ( 直接経費:3600000円 、 間接経費:1080000円 )

    食道癌92症例の手術検体を対象に、活性型のリン酸化SphK1(pSphK1)とSphK1の腫瘍組織における発現を免疫組織化学で評価した。pSphK1高発現群では病理学的なリンパ節転移陽性、リンパ管侵襲陽性、壁内転移陽性の頻度が高かった。さらに、pSphK1高発現群の術後5年全生存割合は50.8%であり、低発現群の67.3%と比較して有意に低かった。一方、SphK1発現とリンパ行性進展や術後生存期間との有意な関連は認められなかった。pSphK1発現は食道癌のリンパ行性進展や患者の不良な予後と関連している。

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  • 覚醒と睡眠の制御におけるセロトニン5HT1A受容体の生理的役割の解明

    研究課題/領域番号:16K21057  2016年4月 - 2018年3月

    日本学術振興会  科学研究費助成事業 若手研究(B)  若手研究(B)

    齊藤 夕貴, 桜井 武, 前島 隆司, 崎村 建司, 阿部 学

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    配分額:3900000円 ( 直接経費:3000000円 、 間接経費:900000円 )

    本研究では5HT1A受容体をオレキシンニューロン特異的に欠損させたox5HT1ARKOマウスを作出し、セロトニンによるオレキシンニューロン制御機構の生理的役割について検討した。5HT1A受容体が欠損しているオレキシンニューロンはセロトニンの抑制を受けず活動レベルを調節することが困難となるため、平常時ではNREM睡眠量が増加することを見出した。また、拘束ストレス負荷後にはオレキシンニューロンの5HT1A受容体の欠損によりREM睡眠量が増加することが分かった。これらの結果からオレキシンニューロンに発現する5HT1A受容体が平常時および拘束ストレス負荷後の睡眠量の調節に関与していることが示唆された。

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  • 記憶と社会的行動に果たす海馬CA2領域の機能解明

    研究課題/領域番号:26350979  2014年4月 - 2017年3月

    日本学術振興会  科学研究費助成事業 基盤研究(C)  基盤研究(C)

    阿部 学

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    配分額:4940000円 ( 直接経費:3800000円 、 間接経費:1140000円 )

    本研究の目的は、機能の不明な部分の多かった海馬CA2領域を介した神経回路の担う記憶や社会的行動に果たす役割を解明することである。標的遺伝子組み換え法によりCA2神経細胞の神経活動のコントロールを可能とするために、DNA組換え酵素Creまたはテトラサイクリントランスアクチベーターを標的遺伝子座(Rgs14、Amigo2、Abatなど)へ挿入したマウスや、マウス生体内におけるCre活性を検証するための新規レポーターマウスを作製し解析を行った。本研究課題において作製された遺伝子改変マウスは、高次脳機能の研究に有用なツールとなることが期待される。

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  • 脳機能解析に特化した全哺乳類胚への応用可能な一世代遺伝子改変動物作成法の開発

    研究課題/領域番号:26640007  2014年4月 - 2016年3月

    日本学術振興会  科学研究費助成事業 挑戦的萌芽研究  挑戦的萌芽研究

    崎村 建司, 阿部 学

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    配分額:3900000円 ( 直接経費:3000000円 、 間接経費:900000円 )

    本研究の目的は、一世代で任意の遺伝子の機能解析を可能とする動物作出技術を開発することである。このためCRISPR/Casシステムを用いたコンディショナルターゲティングに相当する遺伝子改変動物作製法の開発を計画した。
    まず、CreあるいはtTA依存的Cas9発現マウスを樹立し、レンチウィルスベクターによりマウス初期胚へ効率的に外来遺伝子を導入できることを確認した。さらに受精卵を用いた電気穿孔法による低分子タグの一世代ノックインマウス作製法の開発に成功した。これらの技術は原理的に他の哺乳類に適用できることから、サルなどを用いた脳高次機能、疾患の分子機序解析のための基盤技術になると考えられる。

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  • ヒト脳機能解明への道程としての遺伝子改変ラット作製法の開発

    研究課題/領域番号:24240048  2012年4月 - 2016年3月

    日本学術振興会  科学研究費助成事業 基盤研究(A)  基盤研究(A)

    崎村 建司, 阿部 学, 夏目 里恵, 中務 胞, 周 麗

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    配分額:46670000円 ( 直接経費:35900000円 、 間接経費:10770000円 )

    本研究の目的は、脳機能解析に利用できる遺伝子改変ラットを安価かつ容易に作製する方法を開発し、ラットを研究リソースとして利用できる基盤を作ることである。このために、ラットES細胞の培養条件、相同組換えを用いた迅速な遺伝子改変方法、確実に生殖細胞への分化をするキメラ作製法の開発おこない、遺伝子改変ラット樹立技術を確立した。
    さらにラットES細胞を用いて、マウス・ラット異種間キメラ法によりラット精子を作出するために、コンディショナル組換え法により精巣形成不全マウス作出し、高効率でラット精巣組織をマウスで作出することに成功した。本研究により、効率的に遺伝子組換えラットを作る技術が開発された。

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  • コンディショナル遺伝子改変マウスを用いた記憶における海馬CA2領域の機能解析

    研究課題/領域番号:24700321  2012年4月 - 2014年3月

    日本学術振興会  科学研究費助成事業 若手研究(B)  若手研究(B)

    阿部 学

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    配分額:4420000円 ( 直接経費:3400000円 、 間接経費:1020000円 )

    近年まで海馬CA2領域はその機能がほとんど未知であった。本研究の目的はCA2神経細胞の記憶に果たす役割の解明である。そこで、CA2選択的遺伝子改変のためCacng5、Rgs14、AbatなどCA2領域に強く発現することが知られている遺伝子を標的として翻訳開始点にDNA組換え酵素Creまたはテトラサイクリントランスアクチベーターを挿入したノックインマウスや、Cre活性またはテトラサイクリン誘導により神経細胞死を誘導できるマウスを作製した。本研究課題により作製された多数の遺伝子改変マウス系統は、今後、海馬と記憶の研究に極めて有用なツールとなることが期待される。

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  • TDP43の生理機能に注目したALSの病態機序の解明

    研究課題/領域番号:22249036  2010年 - 2012年

    日本学術振興会  科学研究費助成事業 基盤研究(A)  基盤研究(A)

    西澤 正豊, 阿部 学, 桑野 良三, 小野寺 理, 柿田 明美, 佐藤 俊哉, 横関 明男

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    配分額:48880000円 ( 直接経費:37600000円 、 間接経費:11280000円 )

    平成25年6月7日現在我々は,ALSの病態機序としてTDP-43の核から喪失により,核内小体構成蛋白であるSMNの減少が生じ,その結果として生じる核内小体の機能異常が運動神経細胞死に関与している,との仮説をたて,これを検証することを目標とした.そのため,TDP-43の喪失による、培養細胞系,及び患者組織における核内小体の形態について検討した.培養細胞系および患者剖検組織における生化学的解析として,特にTDP-43の減少が核内小体の数に与える影響を検討した.TDP-43を減少させる方法としてはRNAiの手法を用い,培養細胞系にて検討を行った.核内小体のマーカーとして,核内小体であるCajal小体のマーカーとしてcoilin,SMN小体のマーカーとしてSMN, Gem小体のマーカーとしてGeminを対象とし,免疫組織化学法にて解析を加え,これらの数が減少することを確認した.さらにTDP-43変異を有するFALS,さらにSALS患者の残存神経細胞において,同様に核内小体(Cajal小体,SMN小体,Gem小体)の検討を行い,Cajal小体SMN小体,Gem小体が減少していることを見いだした.またGem小体の減少はUsnRNAの減少をきたすことが知られているので,これを検討した.その結果ALS患者での罹患組織ではU12snRNAの減少を認めた.このことからU11/12の成熟障害を示した.

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  • 脳の機能と疾患におけるグルタミン酸受容体の動態

    研究課題/領域番号:21300118  2009年 - 2011年

    日本学術振興会  科学研究費助成事業 基盤研究(B)  基盤研究(B)

    崎村 建司, 夏目 里恵, 阿部 学, 山崎 真弥, 渡辺 雅彦, 狩野 方伸

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    配分額:18850000円 ( 直接経費:14500000円 、 間接経費:4350000円 )

    本研究の目的は、グルタミン酸受容体の発現と安定性、さらにシナプスへの移行と除去が細胞の種類や脳部位により異なった様式で調節され、このことが単純な入力を多様な出力に変換し、複雑な神経機能発現の基礎課程となるという作業仮説を証明することである。この解析ために、4種類のAMPA型グルタミン酸受容体、4種類のNMDA型受容体はじめとして複数のfloxed型標的マウスを樹立した。また、GAD67-Creマウスなど幾つかのCreドライバーマウスを樹立した。これらのマウスを交配させ解析した結果、海馬CA3では、GluN2BがシナプスでのNMDA型受容体の機能発現に必須であることを明らかにした。また、小脳TARPγ-2とγ-7がAMPA型受容体の発現に必須であることを見出した。さらに発達期のシナプスにおいて、GluN2AとGluN2Bが異なった様式でAMPA型受容体を抑制することを単一神経細胞でのノックアウトを用いて示した。

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  • シナプス構成細胞選択的ノックアウトによる小脳皮質シナプス機能制御の分子機構解析

    研究課題/領域番号:21700348  2009年 - 2010年

    日本学術振興会  科学研究費助成事業 若手研究(B)  若手研究(B)

    阿部 学

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    配分額:4290000円 ( 直接経費:3300000円 、 間接経費:990000円 )

    本研究の目的は、グルタミン酸受容体(GluR)複合体の分子動態と小脳皮質シナプス機能制御との関連を明らかにすることである。そのため、シナプスを構成する主要な細胞で選択的にノックアウトできるシステムを開発した。作製された細胞種特異的ノックアウトマウスの解析から、プルキンエ細胞のGluR活性が登上線維及び並行線維の前シナプス機能を調節している可能性や、バーグマングリアに発現するβ-cateninが小脳層構造形成に必須の役割を果たしている可能性を見出すことができた。

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  • C57BL/6マウスにおけるシナプス機能分子の系統的遺伝子改変システムの開発

    研究課題/領域番号:19700336  2007年 - 2008年

    日本学術振興会  科学研究費助成事業 若手研究(B)  若手研究(B)

    阿部 学

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    配分額:3680000円 ( 直接経費:3200000円 、 間接経費:480000円 )

    遺伝子改変マウスの系統的作製のため、BAC-Red/ET システムとMultisite Gateway を組み合わせたポリA トラップ型ターゲティングベクター作製法を実用化し、相同組換え体ES 細胞作製を簡便にする手法を開発した。また、非相同的末端結合関連タンパクの発現抑制により、相同組換え効率を上昇させる手法を確立することができた。開発したこれらの手法を適用する事でシナプス機能遺伝子改変マウスを多数樹立する事ができた。

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  • グルタミン酸受容体のシナプスへの移行と活性調節機序

    研究課題/領域番号:17300099  2005年 - 2007年

    日本学術振興会  科学研究費助成事業 基盤研究(B)  基盤研究(B)

    崎村 建司, 阿部 学, 山崎 真弥

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    配分額:16200000円 ( 直接経費:14700000円 、 間接経費:1500000円 )

    本研究の目的は、興奮性シナプス伝達を担うグルタミン酸受容体のシナプスへの移行と活性調節の分子機序を明らかにすることである。この目的のために、グルタミン酸受容体を構成するサブユニット群を特定の細胞で選択的に欠損できるマウスを系統的に作成し解析をおこなった。標的とした分子は、4種類のAMPA型受容体サブユニットとその受容体の補助サブユニットと我々が考えているTARPγ分子群のうち脳に発現が高いγ2、γ3、γ4、γ5、γ7、γ8である。また、NMDA型受容体では、GluRε1とGluRε2を標的とした。これらの分子をコンディショナルに欠損できるfloxedマウスを、我々が樹立したC57BL/6系統マウスES細胞RENKA株より作成した。TARPγ欠損マウスの解析から、これら分子がいずれもAMPA型受容体のシナプスへの移行と安定性に必須な分子であることが明らかになり、AMPA型受容体の補助サブユニットとして機能していることが証明された。また、終脳に広く発現し、特に海馬興奮性ニューロンに発現の高いTARPγ8欠損マウスを欠損したマウスは、多動と抗不安傾向が認められたので、幾つかの薬剤の影響を検討した。その結果、このマウスは注意欠陥・多動性障害(AD/HD)のモデルになることが明らかになった。一方AMPA型グルタミン酸受容体GluRα1ノックアウトマウスでは、多動と抗不安の表現型は軽微であることから、AD/HDの原因は脳全体でのAMPA型受容体の活性低下ではなく、特定の部位での活性低下であることが示唆された。その責任部位の同定は、AD/HDの病態理解と治療法開発につながる今後に残された課題である。一方、NMDA型受容体GluRε2の海馬CA3ノックアウトマウスの解析から、NMDA型受容体がシナプス領域で活性を持つためにはこのサブユニットが必須な分子であることを明らかにした。

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  • 遺伝子組換え法を用いた神経回路形成・調節機序の分子解析

    研究課題/領域番号:15029217  2003年 - 2004年

    日本学術振興会  科学研究費助成事業 特定領域研究  特定領域研究

    崎村 建司, 阿部 学

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    配分額:6600000円 ( 直接経費:6600000円 )

    本研究の目的は、遺伝子組換え法を用いて神経回路形成・調節機序の分子解析を個体レベルでおこなうことである。この目的のために、脳機能解析に適したC57BL/6系統マウス由来ES細胞から時期・部位特異的に遺伝子欠損が誘導できる手法の開発を進め、新たにES細胞株RENKAを樹立した。この細胞株を用いて、効率よく生殖系列遺伝するキメラマウスが作成できる方法を確立した。また、興奮性シナプス伝達を担うグルタミン酸受容体に着目し、この分子群の機能解析をおこない次のことを明らかにした。生体内でのNMDA型受容体サブユニット構成とシナプスへの移行はその生理機能を理解する上で極めて重要である。そこでGluRε1,GluRε3サブユニットダブル欠失マウスを作成し、NMDA型受容体がシナプスへ移行するにはGluRεサブユニットの存在が必須であり、GluRζ1単独でのシナプスへの移送はほとんど起こらないことを小脳顆粒細胞において明らかにした。さらに、培養細胞での解析から重要性が強調されていたGluRζ1のC末端のスプライシング型は、生体脳におけるこの分子のシナプス表面への移行に影響しないことを示した。また、GluRζ1はGMRεが存在しないと小胞体ですみやかに分解されることが明らかになった。一方、小脳失調症状を呈する突然変異マウスStargazerの原因遺伝子であるstargazin (voltage dependent calcium channel γ2;VDCCγ2)がAMPA型受容体のチャネル活性を直接調節することを見出した。さらに、VDCCγ2以外にもVDCCγ3,γ4,γ7,γ8に同様の活性があることを見出した。これらのことは、AMPA型受容体の活性調節には、VDCCγ分子群の寄与があることを強く示唆する。

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  • コンディショナルターゲッティングを用いた脳高次機能の分子解析

    研究課題/領域番号:13480264  2001年 - 2003年

    日本学術振興会  科学研究費助成事業 基盤研究(B)  基盤研究(B)

    崎村 建司, 阿部 学

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    配分額:15000000円 ( 直接経費:15000000円 )

    遺伝子ノックアウト法は神経系の機能解析に非常に有用な方法であるが、従来のノックアウト法には、いくつかの問題があった。すなわち、1)分子欠損による発生停止・致死、2)分子欠損に由来する形態形成異常(神経ネットワーク等の形成異常)、3)他分子による機能代償などにより、成体でのその分子が持つ正しい機能が解析できなかった。さらに、一般的なノックアウトマウスの作成には129系統マウス由来ES細胞が用いられてきたが、このマウスは脳に奇形があり、脳機能解析には全く向かない。本研究の目的は、これらの問題を解決する方法を開発し、実際にシナプス可塑性に関与する分子の機能を解析することである。我々はC57BL/6系マウス由来のES細胞株を樹立し、この細胞を用いて高効率に生殖系列遺伝するキメラ作成法を実用化することに成功した。この成果は特許申請をおこなった。また、Cre/loxP組換え系を用いてNMDA受容体GluRε2サブユニットおよび細胞接着分子βカテニンのコンディショナルターゲッティングマウスの作成をおこなった。NMDA型グルタミン酸受容体GluRε2サブユニットを海馬CA3領域で欠損したマウスの海馬CA3錐体細胞でのNMDA受容体を介する電流応答を解析した結果、歯状回からの苔状線維、対側のCA3錐体細胞、同側の錐体細胞からそれぞれのシナプスでNMDA受容体を介する電流応答が消失していた。これは、この領域でのNMDA型受容体を介するシナプス伝達にはGluRε2サブユニットが必須であり、GluRε1サブユニットでは代償できない事を示唆する。一方、細胞接着分子βカテニンを海馬CA3領域で欠損したマウスは、正常に成長し交配も可能であったが、文脈依存学習の亢進など行動学的な異常が認められた。今後これら表現型とシナプス伝達効率との関係検証を進める必要がある。

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  • 神経回路の形成、維持、変化における細胞接着分子の役割

    研究課題/領域番号:13041020  2001年 - 2002年

    日本学術振興会  科学研究費助成事業 特定領域研究  特定領域研究

    崎村 建司, 阿部 学

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    配分額:7200000円 ( 直接経費:7200000円 )

    シナプスの形成、維持、変化は、神経系における脳の情報処理の基本的要件であり、これらの分子機序の解明は、脳高次機能を分子レベルで理解するために必須である。本研究の目的は、脳の領域と時期を限定して遺伝子組換えを起こせるシステムを用いて、発達時期のシナプス形成と保持に関与する分子が成体におけるシナプス可塑性や高次脳機能へ果たす役割の検証をおこなう事である。本研究では、脳高次機能を解析する時障害になるマウスの遺伝子背景を避けるために、我々が新たに樹立したC57BL/6系統マウス由来ES細胞を用いた組換えマウス作成システムを利用した。まず、接着分子カドヘリンの裏打ちタンパクであるβカテニン遺伝子に着目した。この分子は、シナプス形成と維持に関与することが示唆されているが、一般的なノックアウトでは胎生14日で発生が停止するために成体での機能が不明である。そこで、この遺伝子にloxP配列を導入した標的マウスを作成し、海馬CA3錐体細胞にCreを優位に発現するγ1CreN+と交配することで、海馬CA3領域でβカテニンを欠損したマウスを作成した。このマウスではβカテニンが海馬CA3領域で欠損していることを免疫組織化学で確認した。さらに、NMDA受容体ε2サブユニットを標的としたマウスとγ1CreN+と交配し、海馬CA3領域でこのサブユニットが発現しないマウスを作成した。NMDA受容体はシナプス可塑性の鍵を握る分子の一つであるがそのε2サブユニットは一般的なノックアウトで致死である。本研究で作成したこれら部位時期特異的ノックアウトマウスは、正常に産まれ生育するために、成体においてそれぞれの分子が果たす機能を解析することが可能である。これらマウスの解析を進め、発達時期のシナプス形成と保持に関与する分子が成体におけるシナプス可塑性や高次脳機能へ果たす役割の検証をおこなう。

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  • 時間空間特異的な遺伝子発現制御系を用いた記憶・学習の分子機構の解明

    研究課題/領域番号:13210056  2001年

    日本学術振興会  科学研究費助成事業 特定領域研究(C)  特定領域研究(C)

    崎村 建司, 阿部 学

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    配分額:7700000円 ( 直接経費:7700000円 )

    シナプス形成過程に関与する分子機構が、シナプスの可塑性発現と記憶・学習にも使用されるという仮説を証明するために、脳の部位と発達段階の時期を限局して機能分子を欠損させたマウスを作成し解析する。このために我々は、学習能力の高いC57BL/6系統マウス由来のES細胞を実用化し、遺伝子背景の問題を解消できるようにした。さらに、活性誘導可能なcreリコンビネースとその認識配列loxPを用いたコンディショナル組換え系を開発した。この系を用いて分子欠損をおこなうために、興奮性シナプス伝達を担うAMPA型グルタミン酸受容体α2サブユニット、カイニン酸型受容体β2、γ1、γ2サブユニットおよびNMDA型受容体ε2サブユニット、シナプス肥厚に局在するPSD95、さらに細胞接着分子βカテニンなどの遺伝子にloxP配列を挿入した標的マウスを作成した。これらのマウスは、FRT-FLP組換え系を用いてオマイシン耐性遺伝子領域を除去し、野生型のマウスと表現型が同一であることを確認した。これらloxP挿入標的マウスと海馬CA3錐体細胞、小脳プルキンエ細胞、小脳穎粒細胞、海馬CA1領域それぞれでcre組換え酵素を発現するマウスを交配させて、部位・時期特異的分子欠損マウスの作製を進めている。これまでに、海馬CA3錐体細胞で特異的にNMDA型受容体ε2サブユニットとβカテニン分子がそれぞれ欠失した個体を得、その表現型を形態学的、電気生理学的、行動学的手法で解析し、シナプス形成過程に関与するこれら分子がシナプスの可塑性発現と記憶・学習にどのような役割を果たすのかを解析している。

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  • コンディショナルジーンターゲティングを用いたシナプス結合形成の分子機構解析

    研究課題/領域番号:98J08474  1998年 - 2000年

    日本学術振興会  科学研究費助成事業 特別研究員奨励費  特別研究員奨励費

    阿部 学

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    配分額:2700000円 ( 直接経費:2700000円 )

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担当経験のある授業科目

  • 統合臨床医学

    2020年
    -
    現在
    機関名:新潟大学

  • 機能生理学

    2014年
    -
    2018年
    機関名:新潟大学