松田 信爾
| 基盤理工学専攻 | 准教授 |
| Ⅲ類（理工系） | 准教授 |
| 脳・医工学研究センター | 准教授 |
- プロフィール:
脳はどのように物事を記憶し学習していくのかを明らかにしたいと考えています。その為に記憶の細胞レベルでの基盤と考えられているシナプス可塑性に分子機構の解明並びにその制御技術の開発を行っています。近年は光を用いて長期抑圧を制御する技術の開発し、小脳長期抑圧が運動学習に直結していることを明らかにしました。長期増強などの他のシナプス可塑性の制御技術も開発し、脳の記憶・学習の実体に迫りたいと考えています。
研究者情報
経歴
- 2014年11月01日
電気通信大学大学院, 情報理工学研究科, 准教授 - 2009年04月01日 - 2014年10月31日
慶応義塾大学, 医学部 生理学教室, 講師 - 2007年04月01日 - 2009年03月31日
慶応義塾大学, 医学部 生理学教室, 助教 - 2003年10月01日 - 2009年03月31日
慶応義塾大学, 医学部 生理学教室, 助手 - 2000年10月01日 - 2003年09月30日
セントジュード小児研究病院, 発達神経科学部門, 博士研究員 - 1998年04月01日 - 2000年09月30日
理化学研究所脳科学総合研究センター, 記憶学習機構研究チーム - 1995年07月01日 - 1998年03月31日
日本学術振興会, 特別研究員ＤＣ１
学歴
研究活動情報
受賞
論文
- Activation of innate immune receptor TLR9 by mitochondrial DNA plays essential roles in the chemical long-term depression of hippocampal neurons
Naoya Atarashi; Misaki Morishita; Shinji Matsuda
ラスト(シニア)オーサー, Journal of Biological Chemistry, Elsevier BV, 300巻, 3号, 掲載ページ 105744-105744, 出版日 2024年03月, 査読付
研究論文（学術雑誌）, 英語 - Subunit-dependent and subunit-independent rules of AMPA receptor trafficking during chemical long-term depression in hippocampal neurons
Shinji Matsuda; Michisuke Yuzaki
筆頭著者, Journal of Biological Chemistry, Cold spring harbor laboratory, 297巻, 2号, 掲載ページ 100949, 出版日 2021年07月10日, 査読付
研究論文（学術雑誌）, 英語 - PIP3-Phldb2 is crucial for LTP regulating synaptic NMDA and AMPA receptor density and PSD95 turnover
Min-Jue Xie; Yasuyuki Ishikawa; Hideshi Yagi; Tokuichi Iguchi; Yuichiro Oka; Kazuki Kuroda; Keiko Iwata; Hiroshi Kiyonari; Shinji Matsuda; Hideo Matsuzaki; Michisuke Yuzaki; Yugo Fukazawa; Makoto Sato
Scientific Reports, Springer Nature Publishing AG, 9巻, 4305号, 掲載ページ 1-15, 出版日 2019年03月13日, 査読付
研究論文（学術雑誌）, 英語 - PhotonSABER: new tool shedding light on endocytosis and learning mechanisms in vivo.
Shinji Matsuda; Wataru Kakegawa; Michisuke Yuzaki
筆頭著者, Communicative & integrative biology, Taylor & Francis, 12巻, 1号, 掲載ページ 34-37, 出版日 2019年, 査読付, 招待, 国際誌, In the central nervous system, activity-dependent endocytosis of postsynaptic AMPA-type glutamate receptors (AMPA receptors) is thought to mediate long-term depression (LTD), which is a synaptic plasticity model in various neuronal circuits. However, whether and how AMPA receptor endocytosis and LTD at specific synapses are causally linked to learning and memory in vivo remains unclear. Recently, we developed a new optogenetic tool, PhotonSABER, which could control AMPA receptor endocytosis in temporal, spatial, and cell-type-specific manners at activated synapses. Using PhotonSABER, we found that AMPA receptor endocytosis and LTD at synapses between parallel fibers and Purkinje cells in the cerebellum mediate oculomotor learning. We also found that PhotonSABER could inhibit endocytosis of epidermal growth factor receptors in HeLa cells upon light stimulation. These results demonstrate that PhotonSABER is a powerful tool for analyzing the physiological functions of endocytosis in non-neuronal cells, as well as the roles of LTD in various brain regions.
研究論文（学術雑誌）, 英語 - エンドサイトーシスを制御する新しい技術により明らかにされた小脳における長期抑圧と運動学習との因果関係
掛川渉; 柚﨑通介; 松田信爾
ライフサイエンス 新着論文レビュー, DOI: 10.7875/first.author.2018巻, 出版日 2018年09月10日, 招待
研究論文（学術雑誌）, 日本語 - Optogenetic Control of Synaptic AMPA Receptor Endocytosis Reveals Roles of LTD in Motor Learning.
Kakegawa W; Katoh A; Narumi S; Miura E; Motohashi J; Takahashi A; Kohda K; Fukazawa Y; Yuzaki M; Matsuda S
ラスト(シニア)オーサー, Neuron, Cell Press, 99巻, 5号, 掲載ページ 985-998, 出版日 2018年09月05日, 査読付, 国際誌, Long-term depression (LTD) of AMPA-type glutamate receptor (AMPA receptor)-mediated synaptic transmission has been proposed as a cellular substrate for learning and memory. Although activity-induced AMPA receptor endocytosis is believed to underlie LTD, it remains largely unclear whether LTD and AMPA receptor endocytosis at specific synapses are causally linked to learning and memory in vivo. Here we developed a new optogenetic tool, termed PhotonSABER, which enabled the temporal, spatial, and cell-type-specific control of AMPA receptor endocytosis at active synapses, while the basal synaptic properties and other forms of synaptic plasticity were unaffected. We found that fiberoptic illumination to Purkinje cells expressing PhotonSABER in vivo inhibited cerebellar motor learning during adaptation of the horizontal optokinetic response and vestibulo-ocular reflex, as well as synaptic AMPA receptor decrease in the flocculus. Our results demonstrate that LTD and AMPA receptor endocytosis at specific neuronal circuits were directly responsible for motor learning in vivo. VIDEO ABSTRACT.
研究論文（学術雑誌）, 英語 - Chemical labelling for visualizing native AMPA receptors in live neurons
Sho Wakayama; Shigeki Kiyonaka; Itaru Arai; Wataru Kakegawa; Shinji Matsuda; Keiji Ibata; Yuri L. Nemoto; Akihiro Kusumi; Michisuke Yuzaki; Itaru Hamachi
NATURE COMMUNICATIONS, NATURE PUBLISHING GROUP, 8巻, 掲載ページ 14850, 出版日 2017年04月, 査読付, The location and number of neurotransmitter receptors are dynamically regulated at postsynaptic sites. However, currently available methods for visualizing receptor trafficking require the introduction of genetically engineered receptors into neurons, which can disrupt the normal functioning and processing of the original receptor. Here we report a powerful method for visualizing native alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA)-type glutamate receptors (AMPARs) which are essential for cognitive functions without any genetic manipulation. This is based on a covalent chemical labelling strategy driven by selective ligand-protein recognition to tether small fluorophores to AMPARs using chemical AMPAR modification (CAM) reagents. The high penetrability of CAM reagents enables visualization of native AMPARs deep in brain tissues without affecting receptor function. Moreover, CAM reagents are used to characterize the diffusion dynamics of endogenous AMPARs in both cultured neurons and hippocampal slices. This method will help clarify the involvement of AMPAR trafficking in various neuropsychiatric and neurodevelopmental disorders.
研究論文（学術雑誌）, 英語 - AP-4
Shinji Matsuda; MIchisuke Yuzaki
Encyclopedia of Signaling Molecules 2nd edition, Springer New York, 掲載ページ 1-6, 出版日 2016年, 招待
研究論文（学術雑誌）, 英語 - Rab8a and Rab8b are essential for several apical transport pathways but insufficient for ciliogenesis
Takashi Sato; Tomohiko Iwano; Masataka Kunii; Shinji Matsuda; Rumiko Mizuguchi; Yongwook Jung; Haruo Hagiwara; Yoshihiro Yoshihara; Michisuke Yuzaki; Reiko Harada; Akihiro Harada
JOURNAL OF CELL SCIENCE, COMPANY OF BIOLOGISTS LTD, 127巻, 2号, 掲載ページ 422-431, 出版日 2014年01月, 査読付, The small GTP-binding protein Rab8 is known to play an essential role in intracellular transport and cilia formation. We have previously demonstrated that Rab8a is required for localising apical markers in various organisms. Rab8a has a closely related isoform, Rab8b. To determine whether Rab8b can compensate for Rab8a, we generated Rab8b-knockout mice. Although the Rab8b-knockout mice did not display an overt phenotype, Rab8a and Rab8b double-knockout mice exhibited mislocalisation of apical markers and died earlier than Rab8a-knockout mice. The apical markers accumulated in three intracellular patterns in the double-knockout mice. However, the localisation of basolateral and/or dendritic markers of the double-knockout mice seemed normal. The morphology and the length of various primary and/or motile cilia, and the frequency of ciliated cells appeared to be identical in control and double-knockout mice. However, an additional knockdown of Rab10 in double-knockout cells greatly reduced the percentage of ciliated cells. Our results highlight the compensatory effect of Rab8a and Rab8b in apical transport, and the complexity of the apical transport process. In addition, neither Rab8a nor Rab8b are required for basolateral and/or dendritic transport. However, simultaneous loss of Rab8a and Rab8b has little effect on ciliogenesis, whereas additional loss of Rab10 greatly affects ciliogenesis.
研究論文（学術雑誌）, 英語 - Stargazin regulates AMPA receptor trafficking through adaptor protein complexes during long-term depression
Shinji Matsuda; Wataru Kakegawa; Timotheus Budisantoso; Toshihiro Nomura; Kazuhisa Kohda; Michisuke Yuzaki
筆頭著者, NATURE COMMUNICATIONS, NATURE PUBLISHING GROUP, 4巻, 掲載ページ 2759, 出版日 2013年11月, 査読付, Long-term depression (LTD) underlies learning and memory in various brain regions. Although postsynaptic AMPA receptor trafficking mediates LTD, its underlying molecular mechanisms remain largely unclear. Here we show that stargazin, a transmembrane AMPA receptor regulatory protein, forms a ternary complex with adaptor proteins AP-2 and AP-3A in hippocampal neurons, depending on its phosphorylation state. Inhibiting the stargazin-AP-2 interaction disrupts NMDA-induced AMPA receptor endocytosis, and inhibiting that of stargazin-AP-3A abrogates the late endosomal/lysosomal trafficking of AMPA receptors, thereby upregulating receptor recycling to the cell surface. Similarly, stargazin's interaction with AP-2 or AP-3A is necessary for low-frequency stimulus-evoked LTD in CA1 hippocampal neurons. Thus, stargazin has a crucial role in NMDA-dependent LTD by regulating two trafficking pathways of AMPA receptors-transport from the cell surface to early endosomes and from early endosomes to late endosomes/lysosomes-through its sequential binding to AP-2 and AP-3A.
研究論文（学術雑誌）, 英語 - The delta 2 glutamate receptor gates long-term depression by coordinating interactions between two AMPA receptor phosphorylation sites
Kazuhisa Kohda; Wataru Kakegawa; Shinji Matsuda; Tadashi Yamamoto; Hisashi Hirano; Michisuke Yuzaki
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, NATL ACAD SCIENCES, 110巻, 10号, 掲載ページ E948-E957, 出版日 2013年03月, 査読付, Long-term depression (LTD) commonly affects learning and memory in various brain regions. Although cerebellar LTD absolutely requires the delta 2 glutamate receptor (GluD2) that is expressed in Purkinje cells, LTD in other brain regions does not; why and how cerebellar LTD is regulated by GluD2 remains unelucidated. Here, we show that the activity-dependent phosphorylation of serine 880 (S880) in GluA2 AMPA receptor subunit, which is an essential step for AMPA receptor endocytosis during LTD induction, was impaired in GluD2-null cerebellum. In contrast, the basal phosphorylation levels of tyrosine 876 (Y876) in GluA2 were increased in GluD2-null cerebellum. An in vitro phosphorylation assay revealed that Y876 phosphorylation inhibited subsequent S880 phosphorylation. Conversely, Y876 dephosphorylation was sufficient to restore S880 phosphorylation and LTD induction in GluD2-null Purkinje cells. Furthermore, megakaryocyte protein tyrosine phosphatase (PTPMEG), which binds to the C terminus of GluD2, directly dephosphorylated Y876. These data indicate that GluD2 gates LTD by coordinating interactions between the two phosphorylation sites of the GluA2.
研究論文（学術雑誌）, 英語 - NMDA Receptor-Mediated PIP5K Activation to Produce PI(4,5)P-2 Is Essential for AMPA Receptor Endocytosis during LTD
Takamitsu Unoki; Shinji Matsuda; Wataru Kakegawa; Ngo Thai Bich Van; Kazuhisa Kohda; Atsushi Suzuki; Yuji Funakoshi; Hiroshi Hasegawa; Michisuke Yuzaki; Yasunori Kanaho
NEURON, CELL PRESS, 73巻, 1号, 掲載ページ 135-148, 出版日 2012年01月, 査読付, NMDA receptor activation leads to clathrin-dependent endocytosis of postsynaptic AMPA receptors. Although this process controls long-term depression (LTD) induction in the hippocampus, how it is regulated by neuronal activities is not completely clear. Here, we show that Ca2+ influx through the NMDA receptor activates calcineurin and protein phosphatase 1 to dephosphorylate phosphatidylinositol 4-phosphate 5-kinase gamma 661 (PIP5K gamma 661), the major phosphatidylinositol 4,5-bisphosphate (PI(4,5)P-2)-producing enzyme in the brain. Bimolecular fluorescence complementation analysis revealed that the dephosphorylated PIP5K gamma 661 became associated with the clathrin adaptor protein complex AP-2 at postsynapses in situ. NMDA-induced AMPA receptor endocytosis and low-frequency stimulation-induced LTD were completely blocked by inhibiting the association between dephosphorylated PIP5K gamma 661 and AP-2 and by overexpression of a kinase-dead PIP5K gamma 661 mutant in hippocampal neurons. Furthermore, knockdown of PIP5K gamma 661 inhibited the NMDA-induced AMPA receptor endocytosis. Therefore, NMDA receptor activation controls AMPA receptor endocytosis during hippocampal LTD by regulating PIP5K gamma 661 activity at postsynapses.
研究論文（学術雑誌）, 英語 - D-Serine regulates cerebellar LTD and motor coordination through the delta 2 glutamate receptor
Wataru Kakegawa; Yurika Miyoshi; Kenji Hamase; Shinji Matsuda; Keiko Matsuda; Kazuhisa Kohda; Kyoichi Emi; Junko Motohashi; Ryuichi Konno; Kiyoshi Zaitsu; Michisuke Yuzaki
NATURE NEUROSCIENCE, NATURE PUBLISHING GROUP, 14巻, 5号, 掲載ページ 603-U93, 出版日 2011年05月, 査読付, D-Serine (D-Ser) is an endogenous co-agonist for NMDA receptors and regulates neurotransmission and synaptic plasticity in the forebrain. D-Ser is also found in the cerebellum during the early postnatal period. Although D-Ser binds to the delta 2 glutamate receptor (GluD2, Grid2) in vitro, its physiological significance has remained unclear. Here we show that D-Ser serves as an endogenous ligand for GluD2 to regulate long-term depression (LTD) at synapses between parallel fibers and Purkinje cells in the immature cerebellum. D-Ser was released mainly from Bergmann glia after the burst stimulation of parallel fibers in immature, but not mature, cerebellum. D-Ser rapidly induced endocytosis of AMPA receptors and mutually occluded LTD in wild-type, but not Grid2-null, Purkinje cells. Moreover, mice expressing mutant GluD2 in which the binding site for D-Ser was disrupted showed impaired LTD and motor dyscoordination during development. These results indicate that glial D-Ser regulates synaptic plasticity and cerebellar functions by interacting with GluD2.
研究論文（学術雑誌）, 英語 - Cerebellar long-term depression requires dephosphorylation of TARP in Purkinje cells.
Toshihiro Nomura; Wataru Kakegawa; Shinji Matsuda; Kazuhisa Kohda; Jun Nishiyama; Takao Takahashi; Michisuke Yuzaki
Eur J Neurosci, 35巻, 掲載ページ 402-410, 出版日 2011年, 査読付
研究論文（学術雑誌）, 英語 - 小脳長期抑制とTARPによる制御(Cerebellar LTD and regulation by TARPs)
野村 寿博; 掛川 渉; 松田 信爾; 幸田 和久; 柚崎 通介
神経化学, (一社)日本神経化学会, 49巻, 2-3号, 掲載ページ 694-694, 出版日 2010年08月
英語 - 細胞内輸送と代謝 AP-4欠損神経細胞軸索におけるAMPA受容体のオートファゴソームへの集積
松田 信爾; 三浦 会理子; 掛川 渉; 幸田 和久; 渡辺 雅彦; 柚崎 通介
解剖学雑誌, (一社)日本解剖学会, 84巻, Suppl.号, 掲載ページ 67-67, 出版日 2009年03月
日本語 - Cbln1 binds to specific postsynaptic sites at parallel fiber-Purkinje cell synapses in the cerebellum
Keiko Matsuda; Tetsuro Kondo; Takatoshi Iijima; Shinji Matsuda; Masahiko Watanabe; Michisuke Yuzaki
EUROPEAN JOURNAL OF NEUROSCIENCE, WILEY-BLACKWELL PUBLISHING, INC, 29巻, 4号, 掲載ページ 707-717, 出版日 2009年02月, 査読付, Cbln1, which belongs to the C1q/tumor necrosis factor superfamily, is a unique molecule that is not only required for maintaining normal parallel fiber (PF)-Purkinje cell synapses, but is also capable of inducing new PF synapses in adult cerebellum. Although Cbln1 is reportedly released from granule cells, where and how Cbln1 binds in the cerebellum has remained largely unclear, partly because Cbln1 undergoes proteolysis to yield various fragments that are differentially detected by different antibodies. To circumvent this problem, we characterized the Cbln1-binding site using recombinant Cbln1. An immunohistochemical analysis revealed that recombinant Cbln1 preferentially bound to PF-Purkinje cell synapses in primary cultures and acute slice preparations in a saturable and replaceable manner. Specific binding was observed for intact Cbln1 that had formed a hexamer, but not for the N-terminal or C-terminal fragments of Cbln1 fused to other proteins. Similarly, mutant Cbln1 that had formed a trimer did not bind to the Purkinje cells. Immunoreactivity for the recombinant Cbln1 was observed in weaver cerebellum (which lacks granule cells) but was absent in pcd cerebellum (which lacks Purkinje cells), suggesting that the binding site was located on the postsynaptic sites of PF-Purkinje cell synapses. Finally, a subcellular fractionation analysis revealed that recombinant Cbln1 bound to the synaptosomal and postsynaptic density fractions. These results indicate that Cbln1, released from granule cells as hexamers, specifically binds to a putative receptor located at the postsynaptic sites of PF-Purkinje cell synapses, where it induces synaptogenesis.
研究論文（学術雑誌）, 英語 - Polarized sorting of AMPA receptors to the somatodendritic domain is regulated by adaptor protein AP-4.
Shinji Matsuda; Mishisuke Yuzaki
Neuroscience Research, 65巻, 掲載ページ 1-5, 出版日 2009年, 査読付
研究論文（学術雑誌）, 英語 - AP-4: Autophagy-four mislocalized proteins in axons
Shinji Matsuda; Michisuke Yuzaki
Autophagy, Taylor and Francis Inc., 4巻, 6号, 掲載ページ 815-816, 出版日 2008年08月16日, 査読付, 招待, Neurons are highly polarized cells composed of two distinct domains, the axon and the somatodendritic domain. Although AMPA-type glutamate receptors, which mediate fast excitatory neurotransmission in the vertebrate CNS, are preferentially expressed in the somatodendritic domain, the molecular mechanisms underlying such polarized distribution have remained elusive. We recently demonstrated that adaptor protein complex-4 (AP-4) binds to transmembrane AMPA receptor regulatory proteins (TARPs), thereby mediating the selective trafficking of AMPA receptors to the somatodendritic domain
generic disruption of AP-4 (AP-4β-/-), results in the mislocalization of TARPs and AMPA receptors in the axons. Similarly, low-density lipoprotein receptors and δ2 glutamate receptors are mislocalized in axons, while other cargos, such as NMDA receptors and metabotropic glutamate receptors, are properly excluded from AP-4β-/- axons. These findings indicate that there exist AP-4-dependent and -independent sorting mechanisms. Unexpectedly, mislocalized AMPA receptors do not reach the cell surface and accumulate in autophagosomes in the bulging portions of AP-4β-/- axons. Several lines of evidence indicate that mislocalized AMPA receptors activate the autophagic pathway. Since increased autophagy and axonal swelling are suggested to occur in various neuronal disorders, further studies using AP-4β-/- mice are warranted to understand the mechanisms regulating autophagy in axons. ©2008 Landes Bioscience.
研究論文（学術雑誌）, 英語 - Accumulation of AMPA receptors in autophagosomes in neuronal axons lacking adaptor protein AP-4
Shinji Matsuda; Eriko Miura; Keiko Matsuda; Wataru Kakegawa; Kazuhisa Kohda; Masahiko Watanabe; Michisuke Yuzaki
NEURON, CELL PRESS, 57巻, 5号, 掲載ページ 730-745, 出版日 2008年03月, 査読付, AP-4 is a member of the adaptor protein complexes, which control vesicular trafficking of membrane proteins. Although AP-4 has been suggested to contribute to basolateral sorting in epithelial cells, its function in neurons is unknown. Here, we show that disruption of the gene encoding the 0 subunit of AP-4 resulted in increased accumulation of axonal autophagosomes, which contained AMPA receptors and transmembrane AMPA receptor regulatory proteins (TARPs), in axons of hippocampal neurons and cerebellar Purkinje cells both in vitro and in vivo. AP-4 indirectly associated with the AMPA receptor via TARPs, and the specific disruption of the interaction between AP-4 and TARPs caused the mislocalization of endogenous AMPA receptors in axons of wild-type neurons. These results indicate that AP-4 may regulate proper somatodendritic-specific distribution of its cargo proteins, including AMPA receptor-TARP complexes and the autophagic pathway in neurons.
研究論文（学術雑誌）, 英語 - Mechanisms for polarized sorting of glutamate receptors to the somatodendritic domain of neurons
Matsuda Shinji; Miura Eriko; Kakegawa Wataru; Matsuda Keiko; Kohda Kazuhisa; Watanabe Masahiko; Yuzaki Michisuke
NEUROSCIENCE RESEARCH, 61巻, 掲載ページ S21, 出版日 2008年, 査読付 - The extreme C-terminus of GluR delta 2 is essential for induction of long-term depression in cerebellar slices
Kazuhisa Kohda; Wataru Kakegawa; Shinji Matsuda; Ryoichi Nakagami; Naomasa Kakiya; Michisuke Yuzaki
EUROPEAN JOURNAL OF NEUROSCIENCE, BLACKWELL PUBLISHING, 25巻, 5号, 掲載ページ 1357-1362, 出版日 2007年03月, 査読付, Long-term depression (LTD) of parallel fibre (PF)-Purkinje cell synapses in the cerebellum is recognized as a cellular substrate of motor learning. Although the delta 2 glutamate receptor (GluR delta 2) has been shown to be crucial for LTD, the mechanisms by which GluR delta 2 functions remain elusive. In this study, we developed a virus vector-based gene transfer approach to rescue impaired LTD in GluR delta 2-null Purkinje cells in cerebellar slice preparations. We demonstrated that LTD was restored in GluR delta 2-null Purkinje cells transduced with wild-type but not with mutant GluR delta 2, which lacked the PDZ-ligand domain in the C-terminus. Immunohistochemical analysis revealed no difference in expression levels or spine localization patterns between virally introduced wild-type and mutant GluR delta 2 proteins. Similarly, LTD was abrogated in Purkinje cells that had been acutely perfused with peptides, hampering the interaction of GluR delta 2 with PDZ proteins such as PSD-93, PTPMEG and S-SCAM but not with delphilin. Together, these results indicate that PDZ proteins that bind to the C-terminus of GluR delta 2 are not essential for localizing GluR delta 2 at synapses but are crucial for conveying signals necessary for the induction of LTD.
研究論文（学術雑誌）, 英語 - Characterization of the delta 2 glutamate receptor-binding protein delphilin - Splicing variants with differential palmitoylation and an additional PDZ domain
Keiko Matsuda; Shinji Matsuda; Clare M. Gladding; Michisuke Yuzaki
JOURNAL OF BIOLOGICAL CHEMISTRY, AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC, 281巻, 35号, 掲載ページ 25577-25587, 出版日 2006年09月, 査読付, The glutamate receptor delta 2 (GluR delta 2) is predominantly expressed at parallel fiber-Purkinje cell postsynapses and plays crucial roles in synaptogenesis and synaptic plasticity. Although the mechanism by which GluR delta 2 functions remains unclear, its lack of channel activity and its role in controlling the endocytosis of alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionate ( AMPA) receptors have suggested that GluR delta 2 may convey signals by interacting with intracellular signaling molecules. Among several proteins that interact with GluR delta 2, delphilin is unique in that it is selectively expressed at parallel fiber-Purkinje cell synapses and that, in addition to a single PDZ domain, it contains a formin homology domain that is thought to regulate actin dynamics. Here, we report a new isoform of delphilin, designated as L-delphilin, that has alternatively spliced N-terminal exons encoding an additional PDZ domain. Although original delphilin, designated S-delphilin, was palmitoylated at the N terminus, this region was spliced out in L-delphilin. As a result, S-delphilin was associated with plasma membranes in COS cells and dendritic spines in hippocampal neurons, whereas L-delphilin formed clusters in soma and dendritic shafts. In addition, S-delphilin, but not L-delphilin, facilitated the expression of GluR delta 2 on the cell surface. These results indicate that, like PSD-95 and GRIP/ABP, delphilin isoforms with differential palmitoylation and clustering capabilities may provide two separate intracellular and surface GluR delta 2 pools and may control GluR delta 2 signaling in Purkinje cells.
研究論文（学術雑誌）, 英語 - A new motif necessary and sufficient for stable localization of the delta 2 glutamate receptors at postsynaptic spines
S Matsuda; K Matsuda; M Yuzaki
JOURNAL OF BIOLOGICAL CHEMISTRY, AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC, 281巻, 25号, 掲載ページ 17501-17509, 出版日 2006年06月, 査読付, The number of each subclass of ionotropic glutamate receptors (iGluRs) at the spines is differentially regulated either constitutively or in a neuronal activity-dependent manner. The delta 2 glutamate receptor (GluR delta 2) is abundantly expressed at the spines of Purkinje cell dendrites and controls synaptic plasticity in the cerebellum. To obtain clues to the trafficking mechanism of the iGluRs, we expressed wildtype or mutant GluR delta 2 in cultured hippocampal and Purkinje neurons and analyzed their intracellular localization using immunocytochemical techniques. Quantitative analysis revealed that deletion of the 20 amino acids at the center of the C terminus (region E) significantly reduced the amount of GluR delta 2 protein at the spines in both types of neurons. This effect was partially antagonized by the inhibition of endocytosis by high dose sucrose treatment or coexpression of dominant negative dynamin. In addition, mutant GluR delta 2 lacking the E region (GluR delta 2(Delta E)), but not wild-type GluR delta 2, was found to colocalize with the endosomal markers Rab4 and Rab7. Moreover, the antibody-feeding assay revealed that GluR delta 2(Delta E) was internalized more rapidly than GluR delta 2(wt). These results indicate that the E region (more specifically, a 12-amino-acid-long segment of the E2 region) is necessary for rendering GluR delta 2 resistant to endocytosis from the cell surface at the spines. Furthermore, insertion of the E2 region alone into the C terminus of the GluR1 subtype of iGluRs was sufficient to increase the amount of GluR1 proteins in the spines. Therefore, we propose that the E2 region of GluR delta 2 is necessary, and also sufficient, to inhibit endocytosis of the receptor from postsynaptic membranes.
研究論文（学術雑誌）, 英語 - Roles of the N-terminal domain on the function and quaternary structure of the ionotropic glutamate receptor
S Matsuda; Y Kamiya; M Yuzaki
JOURNAL OF BIOLOGICAL CHEMISTRY, AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC, 280巻, 20号, 掲載ページ 20021-20029, 出版日 2005年05月, 査読付, The α-amino-3-hydroxyl-5-methyl-4-isoxazolepropionic acid (AMPA) subtype of ionotropic glutamate receptors (iGluRs) mediates fast excitatory neurotransmission in the mammalian brain. Although the most N-terminal leucine/isoleucine/valine-binding protein (LIVBP) domain is suggested to play a role in the initial assembly of iGluR subunits, it is unclear how this domain is arranged and functions in intact iGluRs. Similarly, although recent crystallographic analyses indicate that the isolated ligand-binding lysine/arginine/ornithine-binding protein domain forms a 2-fold symmetric dimer, the subunit stoichiometry of intact iGluRs remains elusive. Here, we developed a new approach to address these issues. The LIVBP domain of the GluR1 subunit of AMPA receptors was replaced by leucine-zipper peptides designed to form stable symmetric dimers, trimers, tetramers, or pentamers. All these mutant GluR1s were expressed in human embryonic kidney 293 cells and were transported to the cell surface as well as wild type GluR1. Functional and biochemical analyses indicated that these oligomerizing peptides specifically controlled the formation of the expected number of subunits in a channel complex. However, the channel function was only restored by the tetramer-forming peptide. Although the purified LIVBP domain of GluR1 formed a dimmer in solution, a dimer-forming peptide could not restore the function of GluR1. Moreover, a cross-linking assay indicated that four LIVBP domains are located in proximity to each other. These results suggest that the function of the LIVBP domain is not simply to form initial dimers but to adopt a conformation compatible with the overall tetrameric arrangement of subunits in intact AMPA receptors.
研究論文（学術雑誌）, 英語 - Rescue of abnormal phenotypes of the delta 2 glutamate receptor-null mice by mutant delta 2 transgenes
H Hirai; T Miyazaki; W Kakegawa; S Matsuda; M Mushina; M Watanabe; M Yuzaki
EMBO REPORTS, NATURE PUBLISHING GROUP, 6巻, 1号, 掲載ページ 90-95, 出版日 2005年01月, 査読付, The delta2 glutamate receptor (GluRdelta2) has a crucial role in cerebellar functions; disruption of GluRdelta2 alleles in mice (delta2(-/-)) impairs synapse formation and long-term depression, which is thought to underlie motor learning in the cerebellum, and consequently leads to motor discoordination. However, it has been unclear whether GluRdelta2 is activated by glutamate analogues. Here we introduced a GluRdelta2 transgene, which had a mutation (Arg514Lys) in the putative ligand-binding motif conserved in all mammalian ionotropic glutamate receptors (iGluRs) and their ancestral bacterial periplasmic amino-acid-binding proteins, into delta2(-/-) mice. Surprisingly, a mutant GluRdelta2 transgene, as well as a wild-type GluRdelta2 transgene, rescued all abnormal phenotypes of delta2(-/-) mice. Therefore, these results indicate that the conserved arginine residue, which is crucial for the binding of iGluRs to glutamate analogues, is not essential for the restoration of GluRdelta2 functions in delta2(-/-) mice.
研究論文（学術雑誌）, 英語 - The C-terminal juxtamembrane region of the delta 2 glutamate receptor controls its export from the endoplasmic reticulum
S Matsuda; R Hannen; K Matsuda; N Yamada; T Tubbs; M Yuzaki
EUROPEAN JOURNAL OF NEUROSCIENCE, WILEY-BLACKWELL, 19巻, 7号, 掲載ページ 1683-1690, 出版日 2004年04月, 査読付, Functions of ionotropic glutamate receptors (iGluRs) are tightly regulated by the intracellular trafficking of receptor proteins. Unlike other iGluRs that are considerably retained in the intracellular component, the delta2 glutamate receptor (GluRdelta2) is efficiently expressed on the Purkinje cell surface. To understand the trafficking mechanism of iGluRs, we deleted various portions of the C-terminal intracellular domain of GluRdelta2 and analysed the localization of the mutant proteins in heterologous cells and neurons. Biotinylation assays indicated that GluRdelta2 lacking the C-terminal juxtamembrane region of 13 amino acids (region A) was not present on the cell surface. This mutant GluRdelta2 was sensitive to endoglycosidase H, which digests unprocessed high-mannose oligosaccharides on proteins retained in the endoplasmic reticulum (ER) or cis-Golgi. Therefore, we concluded that region A is crucial for the transport of GluRdelta2 beyond the trans-Golgi to the cell surface. Because the immunostaining pattern of GluRdelta2 lacking region A in cultured hippocampal neurons completely overlapped the pattern of fluorescence emitted by ER-resident green fluorescent protein, region A is most likely necessary for GluRdelta2's exit from the ER. Furthermore, this region is essential for the proper intracellular trafficking of GluRdelta2 in Purkinje cells. Region A does not rely on a dihydrophobic motif or positively charged residues to participate in trafficking, but its function is dependent on the juxtamembrane position. Therefore, we propose that GluRdelta2's efficient transport to the cell surface utilizes an unknown but general ER exit mechanism, which probably works in close relation to the membrane of heterologous cells and neurons.
研究論文（学術雑誌）, 英語 - A hot spot for hotfoot mutations in the gene encoding the delta 2 glutamate receptor
Y Wang; S Matsuda; Drews, V; T Torashima; MH Meisler; M Yuzaki
EUROPEAN JOURNAL OF NEUROSCIENCE, WILEY-BLACKWELL, 17巻, 8号, 掲載ページ 1581-1590, 出版日 2003年04月, 査読付, The orphan glutamate receptor delta2 is selectively expressed in Purkinje cells and plays a crucial role in cerebellar functions. Recently, ataxia in the hotfoot mouse ho4J was demonstrated to be caused by a deletion in the delta2 receptor gene (Grid2 ) removing the N-terminal 170 amino acids of the delta2 receptor. To understand how delta2 receptors function, we characterized mutations in eight additional spontaneously occurring hotfoot alleles of Grid2 . The mouse Grid2 gene consists of 16 exons, spanning approximately 1.4 Mb. Genomic DNA analysis showed that seven hotfoot mutants had a deletion of one or more exons encoding the N-terminal domain of delta2 receptors. The exception is ho5J , which has a point mutation in exon 12. Deletions in ho7J, ho9J , ho11J and ho12J mice result in the in-frame deletion of between 40 and 95 amino acids. Expression of constructs containing these deletions in HEK293 cells resulted in protein retention in the endoplasmic reticulum or cis -Golgi without transport to the cell surface. Coimmunoprecipitation assays indicated that these deletions also reduce the intermolecular interaction between individual delta2 receptors. These results indicate that the deleted N-terminal regions are crucial for oligomerization of delta2 receptors and their subsequent transport to the cell surface of Purkinje cells. The relatively large size of the Grid2 gene may be one of the reasons why many spontaneous mutations occur in this gene. In addition, the frequent occurrence of in-frame deletions within the N-terminal domain in hotfoot mutants suggests the importance of this domain in the function of delta2 receptors.
研究論文（学術雑誌）, 英語 - Heteromer formation of delta 2 glutamate receptors with AMPA or kainate receptors
K Kohda; Y Kamiya; S Matsuda; K Kato; H Umemori; M Yuzaki
MOLECULAR BRAIN RESEARCH, ELSEVIER SCIENCE BV, 110巻, 1号, 掲載ページ 27-37, 出版日 2003年01月, 査読付, The 82 glutamate receptor (GluRdelta2) is predominantly expressed in the postsynaptic densities of parallel fiber-Purkinje cell synapses and plays a crucial role in cerebellar function. However, the mechanisms by which GluRdelta2 participates in cerebellar functions are largely unknown because GluRdelta2 does not bind glutamate analogs. We investigated the possibility that GluRdelta2 may be involved in channel formation together with other glutamate receptor families. We transiently expressed lurcher mutant AMPA receptor GluR1(Lc) and kainate receptor GluR6(Lc) in HEK293 cells. Cells expressing these constitutively active channels displayed a rectifying current-voltage (I-V) relationship. However, when cells were co-transfected with GluRdelta2(Lc), which had the arginine residue in the channel pore region, cells displayed a linear I-V relationship, a result that indicates GluRdelta2(Lc) formed functional heteromeric channels with GluR1(Lc) or GluR6(Lc). Assembly of GluRdelta2 with GluR1 or GluR6 was further confirmed by co-immunoprecipitation assays in HEK293 cells. In addition, GluRdelta2 receptors were partially co-immunoprecipitated from cerebellar synaptosomal fractions by antibodies against GluR2 or KA2. In contrast to lurcher channels. expression of wild-type GluRdelta2 significantly reduced the glutamate-induced current of the wild-type GluR1 receptors without affecting channel properties, such as current kinetics, dose-response relationship, and single-channel conductance. Thus, the heteromeric channel created by the association of wild-type GluR1 and GluRdelta2 may not be gated by glutamate and does not participate in glutamate-induced currents. These results suggest that GluRdelta2 and AMPA or kainate receptors can assemble to form heteromeric receptors in vitro and could modify glutamate signaling in vivo. These findings may help explain the role of GluRdelta2. (C) 2002 Elsevier Science B.V. All rights reserved.
研究論文（学術雑誌）, 英語 - Mutation in hotfoot-4J mice results in retention of delta 2 glutamate receptors in ER
S Matsuda; M Yuzaki
EUROPEAN JOURNAL OF NEUROSCIENCE, WILEY-BLACKWELL, 16巻, 8号, 掲載ページ 1507-1516, 出版日 2002年10月, 査読付, The orphan glutamate receptor delta2 is selectively expressed in Purkinje cells and plays a critical role in cerebellar function. Recently, the ataxia of hotfoot-4J (ho-4J) mice was shown to be caused by a 170-amino acid deletion in the N-terminal region of delta2 receptors. To understand delta2 receptor function, we characterized these mutant receptors (delta2(ho)) in Purkinje cells. Immunohistochemical staining showed that delta2(ho) receptors of the ho-4J homozygotes were abundantly expressed but localized to the Purkinje cell soma; in wild-type mice, delta2 receptors were predominantly present at distal dendrites of Purkinje cells. In addition, delta2(ho) receptors of the ho-4J mice were sensitive to endoglycosidase H, a finding suggesting that delta2(ho) receptors were not transported beyond the endoplasmic reticulum (ER) or cis-Golgi apparatus. To gain further insights into the mechanisms of this phenomenon, we characterized delta2(ho) receptors in transfected HEK293 cells. delta2(ho) receptors expressed in HEK293 cells were also sensitive to endoglycosidase H. Immunohistochemical staining showed that delta2(ho) receptors colocalized with proteins retained in the ER. Furthermore, delta2(ho) receptors were not labelled by membrane-impermeable biotinylation reagents. Coimmunoprecipitation assays showed that the intermolecular interaction of delta2(ho) receptors was significantly weaker than those of wild-type delta2 receptors, a finding suggesting that the ho-4J region is involved in oligomerization of delta2 receptors. Thus, delta2(ho) receptors were retained in the ER, probably by the quality control mechanism that detects unstable oligomers. We conclude that the absence of delta2 receptors on the cell surface by failed transport from the ER of Purkinje cells causes ataxia.
研究論文（学術雑誌）, 英語 - Cloning and characterization of a novel NMDA receptor subunit NR3B: a dominant subunit that reduces calcium permeability.
Keiko Matsuda; Yoshinori Kamiya; Shinji Matsuda; Michisuke Yuzaki
Brain Res Mol Brain Res, 100巻, 掲載ページ 43-52, 出版日 2002年, 査読付
研究論文（学術雑誌）, 英語 - Disruption of AMPA receptor GluR2 clusters following long-term depression induction in cerebellar Purkinje neurons
S Matsuda; T Launey; S Mikawa; H Hirai
EMBO JOURNAL, OXFORD UNIV PRESS, 19巻, 12号, 掲載ページ 2765-2774, 出版日 2000年06月, 査読付, Cerebellar long-term depression (LTD) is thought to play an important role in certain types of motor learning. However, the molecular mechanisms underlying this event have not been clarified. Here, using cultured Purkinje cells, we show that stimulations inducing cerebellar LTD cause phosphorylation of Ser880 in the intracellular C-terminal domain of the AMPA receptor subunit GluR2. This phosphorylation is accompanied by both a reduction in the affinity of GluR2 to glutamate receptor interacting protein (GRIP), a molecule known to be critical for AMPA receptor clustering, and a significant disruption of postsynaptic GluR2 clusters. Moreover, GluR2 protein released from GRIP is shown to be internalized. These results suggest that the dissociation of postsynaptic GluR2 clusters and subsequent internalization of the receptor protein, initiated by the phosphorylation of Ser880, are the mechanisms underlying the induction of cerebellar LTD.
研究論文（学術雑誌）, 英語 - Phosphorylation of serine-880 in GluR2 by protein kinase C prevents its C terminus from binding with glutamate receptor-interacting protein
S Matsuda; S Mikawa; H Hirai
JOURNAL OF NEUROCHEMISTRY, LIPPINCOTT WILLIAMS & WILKINS, 73巻, 4号, 掲載ページ 1765-1768, 出版日 1999年10月, 査読付, Phosphorylation of the glutamate receptor is an important mechanism of synaptic plasticity. Here, we show that the C terminus of GluR2 of the alpha-amino-3-hydroxy-5-methylisoxazole-4-propionate (AMPA) receptor is phosphorylated by protein kinase C and that serine-880 is the major phosphorylation site. This phosphorylation also occurs in human embryonic kidney (HEK) cells by addition of 12-O-tetradecanoylphorbol 13-acetate. Our immunoprecipitation experiment revealed that the phosphorylation of serine-880 in GluR2 drastically reduced the affinity for glutamate receptor-interacting protein (GRIP), a synaptic PDZ domain-containing protein, in vitro and in HEK cells. This result suggests that modulation of serine-880 phosphorylation in GluR2 controls the clustering of AMPA receptors at excitatory synapses and consequently contributes to synaptic plasticity.
研究論文（学術雑誌）, 英語 - Interaction of the C-terminal domain of delta glutamate receptor with spectrin in the dendritic spines of cultured Purkinje cells
H Hirai; S Matsuda
NEUROSCIENCE RESEARCH, ELSEVIER SCI IRELAND LTD, 34巻, 4号, 掲載ページ 281-287, 出版日 1999年09月, 査読付, The interaction of neurotransmitter receptors with the underlying cytoskeleton via subsynaptic proteins is an important mechanism for the targeting of the receptors to synapses in the central nervous system. We show that delta glutamate receptors (delta receptors), expressed predominantly in the dendritic spines of cerebellar Purkinje cells, directly interact with spectrin, a member of the actin-binding family of proteins. Moreover, the interaction between spectrin and C-terminal domain of the delta receptor is 50% inhibited by 1 mu M of Ca2+ in vitro, compared with that in the absence of Ca2+. These results suggest that delta receptors on the postsynaptic membrane of the dendritic spines of cerebellar Purkinje cells are anchored to the actin cytoskeleton via spectrin, and that Ca2+ elevation in the dendritic spines causes delta receptor declustering by dissociation of the receptors from spectrin. This mechanism for receptor anchoring at postsynaptic sites may regulate synaptogenesis and/or synaptic plasticity. (C) 1999 Elsevier Science Ireland Ltd. All rights reserved.
研究論文（学術雑誌）, 英語 - The clustering of NMDA receptor NR1 subunit is regulated by the interaction between the C-terminal exon cassettes and the cytoskeleton
S Matsuda; H Hirai
NEUROSCIENCE RESEARCH, ELSEVIER SCI IRELAND LTD, 34巻, 3号, 掲載ページ 157-163, 出版日 1999年08月, 査読付, The clustering of neurotransmitter receptors at postsynaptic sites is considered to play an important role in modulating synaptic efficacy. To investigate the mechanisms underlying neurotransmitter receptor clustering, we expressed the NMDA (N-methyl-D-aspartate) receptor NR1A subunit in human embryonic kidney (HEK) 293 cells. As previously shown, the cells exhibited subcellular clusters of the receptor protein with a mean diameter of approximately 0.7 mu m. To examine the involvement of cytoskeletal structures on this clustering, we disrupted actin filaments or microtubules by treating the cells with alkaloids. In the actin filament-disrupted cells. the receptor protein shifted from the cellular membrane to the cytoplasm where it formed macroclusters (approximate diameter 3 mu m). In the microtubule-disrupted cells, the subcellular clusters of NR1A could not be detected and the protein was diffusely distributed throughout the cytoplasm. Similar results were obtained by coexpression of the receptor protein with fusion proteins harboring various C-terminal exon cassettes. These results suggest that subcellular clustering of the NR1 subunit of the NMDA receptors is regulated by the interaction of its C-terminus with cytoskeletal components, where differentially spliced cassettes interact separately with actin filaments or microtubules. Modulation of the interaction between the neurotransmitter receptors and the cytoskeleton leads to the rearrangement of the receptor clusters and may contribute to certain types of synaptic plasticity. (C) 1999 Elsevier Science Ireland Ltd. All rights reserved.
研究論文（学術雑誌）, 英語 - Role of carboxyl-terminal charges on S-modulin membrane affinity and inhibition of rhodopsin phosphorylation
S Matsuda; O Hisatomi; F Tokunaga
BIOCHEMISTRY, AMER CHEMICAL SOC, 38巻, 4号, 掲載ページ 1310-1315, 出版日 1999年01月, 査読付, S-Modulin shows a higher affinity for urea-stripped frog rod outer segment membranes than s26 (a cone homologue of S-modulin). NaCl at a concentration of several hundred millimolar reduced the membrane affinity of S-modulin to the s26 level. Chimeric S-modulin and s26 whose respective 23 and 29 amino acids at the carboxyl terminus were swapped showed membrane affinites similar to those of s26 and S-modulin, respectively. The membrane affinity of an S-modulin mutant lacking C-terminal positive charges was reduced to the s26 level, while another S-modulin mutant lacking C-terminal negative charges has a higher membrane affinity than wild-type S-modulin. When the molar ratio of recombinant S-modulins To rhodopsin is 0.5, there was no large difference in the inhibition efficiency. However, S-modulin and mutants with high membrane affinities inhibit rhodopsin phosphorylation more efficiently than s26 and mutants with low membrane affinities at the molar ratio of 0.1. These results indicate that the C-terminal positive charges of these Ca2+-binding proteins enhance the membrane affinity and the inhibitory effect on rhodopsin phosphorylation by increasing the concentration of S-modulin on the membrane.
研究論文（学術雑誌）, 英語 - Evolution of visual pigments and related molecules
Fumio Tokunaga; Osamu Hisatomi; Takunori Satoh; Yuki Taniguchi; Shinji Matsuda; Yoshikazu Imanishi; Hanayo Honkawa; Yusuke Takahashi; Yuko Kobayashi; Masao Yoshida; Yasuo Tsukahara
Novartis Foundation Symposium, 224巻, 掲載ページ 44-53, 出版日 1999年, 査読付, The molecular phylogenetic tree of vertebrate visual pigments, constructed on the basis of amino acid sequence identity, suggests that the visual pigments can be classified into five groups (L, ML, MS, S and Rh) and that their genes have evolved along these five gene lines. Goldfish has a UV-sensitive visual pigment (S group) localized in miniature single cone cells. Medaka has one type of rod cell containing rhodopsin (Rh group) and four types of cone cells, each of which contains a specific visual pigment with an absorption maximum that differs from those of the others. Frogs have a violet-sensitive visual pigment (S group) in small single cone cells and a blue-sensitive visual pigment (MS group) in green rod cells. Although nocturnal and diurnal geckos have rod- and cone-based retinas, respectively, they have phylogenetically closely related visual pigments. The pigments in each line may have restricted absorption maxima. We have cloned cDNAs encoding molecules involved in the phototransduction system of visual cells, such as phosphodiesterase, opsin kinase and arrestin. We then constructed phylogenetic trees of these molecules with the deduced amino acid sequences. The resulting phylogenetic trees show that these molecules are classified into two groups
one is expressed in cones and another in rods, suggesting that rods and cones contain homologous molecules with different amino acid sequences. These differences may result in the different light responses of rods and cones.
研究論文（学術雑誌）, 英語 - The role of calcium-binding sites in S-modulin function
S Matsuda; O Hisatomi; T Ishino; Y Kobayashi; F Tokunagati
JOURNAL OF BIOLOGICAL CHEMISTRY, AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC, 273巻, 32号, 掲載ページ 20223-20227, 出版日 1998年08月, 査読付, S-modulin controls rhodopsin phosphorylation in a calcium-dependent manner, and it has been suggested that it modulates the light sensitivity of the photoreceptor cell, S-modulin binds to the ROS membrane at high Ca2+ concentration, and N-terminal myristoylation is necessary for this property (the calcium-myristoyl switch). S-modulin has four EF-hand motifs, of which two (EF-2 and -3) are functional. Here, we report on the roles of EF-2 and -3 in S-modulin function (calcium binding, membrane association, and inhibition of rhodopsin phosphorylation) by site-directed mutants (E85M and E121M), Surprisingly, E121M, which has a mutation in EF-3, neither binds Ca2+ nor inhibits phosphorylation, In contrast, E85M binds one Ca2+ and has the same membrane affinity as wild-type S-modulin, but has lost the ability to inhibit rhodopsin phosphorylation, It is suggested that the binding of Ca2+ to EF-3 is probably required for EF-2 to be a functional Ca2+-binding site and to induce exposure of the myristoyl group; and that the binding of Ca2+ to EF-2 is important for the interaction with rhodopsin kinase.
研究論文（学術雑誌）, 英語 - A novel subtype of G-protein-coupled receptor kinase, GRK7, in teleost cone photoreceptors
O Hisatomi; S Matsuda; T Satoh; S Kotaka; Y Imanishi; F Tokunaga
FEBS LETTERS, ELSEVIER SCIENCE BV, 424巻, 3号, 掲載ページ 159-164, 出版日 1998年03月, 査読付, Two kinds of retinal cDNA fragments (OlGRK-R and -C) encoding the putative G-protein-coupled receptor kinases (GRKs) were isolated from medaka, Oryzias latipes, OlGRK-R appears to be closely related to the rhodopsin kinase (RK) found in the outer segments of mammalian photoreceptors, but the deduced amino acid sequence of OlGRK-C shows less than 50% identity to those of GRKs known to date, suggesting that OlGRK-C is a novel GRK subtype (GRK7). The mRNA of OlGRK-R is detectable in rods, and that of OlGRK-C is found in ail four types of cone photoreceptor. The C-terminal of OlGRK-R has a consensus sequence for farnesylation, whereas, surprisingly, OlGRK-C has a consensus sequence for geranylgeranylation, Our result are consistent with the concept that lower vertebrates have rod-and cone-specific opsin kinases. (C) 1998 Federation of European Biochemical Societies.
研究論文（学術雑誌）, 英語 - Functional expression and characterization of frog photoreceptor-specific calcium-binding proteins
O Hisatomi; T Ishino; S Matsuda; K Yamaguchi; Y Kobayashi; S Kawamura; F Tokunaga
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, ACADEMIC PRESS INC ELSEVIER SCIENCE, 234巻, 1号, 掲載ページ 173-177, 出版日 1997年05月, 査読付, S-modulin (sensitivity-modulating protein) is a photoreceptor-specific calcium-binding protein which plays an important role in the light adaptation process by controlling rhodopsin phosphorylation in rods. S-modulin and its cone homologue, s26, were expressed at high level (more than 30% of total protein) in Escherichia coli and then purified. They both inhibited rhodopsin phosphorylation in a calcium dependent manner. Myristoylated recombinants of S-modulin and s26 showed calcium-dependent changes in tryptophan emission spectra with half-maxima at about 0.7 mu M free calcium concentration. However, the spectral changes are distinctive from each other, suggesting that there is some difference in the structural change between S-modulin and s26. (C) 1997 Academic Press.
研究論文（学術雑誌）, 英語 - Photoreceptor protein s26, a cone homologue of S-modulin in frog retina
S Kawamura; O Kuwata; M Yamada; S Matsuda; O Hisatomi; F Tokunaga
JOURNAL OF BIOLOGICAL CHEMISTRY, AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC, 271巻, 35号, 掲載ページ 21359-21364, 出版日 1996年08月, 査読付, A frog retinal protein named s26 is a 26-kDa protein found during purification of S-modulin in frog retina (Kawamura, S. (1992) Photochem. Photobiol. 56, 1173-1180). To identify its role in frog retina, first s26 was purified to nearly homogeneity with three chromatographical steps, Based on the partial amino acid sequences of the proteolysed fragments of s26, we isolated cDNAs that encode s26. The analysis of its amino acid sequence revealed that s26 is an S-modulin-like protein, while it shows higher homology to visinin. Visinin is a Ca2+-binding protein reported to be present in chicken cones, but its localization in the retina had been a subject in dispute, The present study showed that s26 is present in cone photoreceptors, The study also showed that s26 inhibits phosphorylation of rhodopsin after a light flash at high Ca2+ concentrations as S-modulin does. From these results, we concluded that s26 is a cone homologue of S-modulin, The result is consistent with the idea that each type of photoreceptors expresses each cell-type specific version of phototransduction proteins.
研究論文（学術雑誌）, 英語
MISC
- Chemical LTD stimulation induced the drastic mitochondrial morphological changes in cultured hippocampal neurons
Naoya Atarashi; Shinji Matsuda
ラスト(シニア)オーサー, 出版日 2023年03月, The Journal of Physiological Sciences, 73巻, 11号, 掲載ページ 130-130, 英語, 研究発表ペーパー・要旨（全国大会，その他学術会議） - Mechanism of hippocampal adult-born neurons for memory consolidation
Iyo Koyanagi; Yuteng Wang; Sakthivel Srinivasan; Jiahui Yu; Toshie Naoi; Akinobu Ohba; Deependra Kumar; Pei-Hsi Wu; Shinji Matsuda; Yoan Cherasse; Takeshi Sakurai; Masanori Yanagisawa; Kaspar Vog; Michisuke Yuzaki; Masanori Sakaguti
出版日 2022年12月, The Journal of Physiological Sciences, 72巻, 33号, 掲載ページ 151-151, 英語, 研究発表ペーパー・要旨（全国大会，その他学術会議） - 【小脳学習説Marr-Albus-Ito理論の50年】小脳学習における長期抑圧(LTD)の役割
掛川 渉; 松田 信爾; 柚崎 通介
(株)中外医学社, 出版日 2019年08月, Clinical Neuroscience, 37巻, 8号, 掲載ページ 919-924, 日本語, 0289-0585, 2020049431 - Optogenetic control of the function of the Golgi apparatus in neurons
Junnosuke Endo; Yukari Takeo; Shinji Matsuda; Michisuke Yuzaki
OXFORD UNIV PRESS INC, 出版日 2014年11月, GLYCOBIOLOGY, 24巻, 11号, 掲載ページ 1201-1201, 英語, 研究発表ペーパー・要旨（国際会議）, 0959-6658, 1460-2423, WOS:000344670300322 - Stargazin regulates AMPA receptor trafficking from plasma membrane to early endosome and lysosome during long term depression
Shinji Matsuda; Michisuke Yuzaki
SPRINGER JAPAN KK, 出版日 2013年, JOURNAL OF PHYSIOLOGICAL SCIENCES, 63巻, 掲載ページ S28-S28, 英語, 研究発表ペーパー・要旨（国際会議）, 1880-6546, WOS:000322352200045 - How does the delta 2 glutamate receptor regulate cerebellar LTD?
Kazuhisa Kohda; Wataru Kakegawa; Shinji Matsuda
SPRINGER JAPAN KK, 出版日 2013年, JOURNAL OF PHYSIOLOGICAL SCIENCES, 63巻, 掲載ページ S38-S38, 英語, 研究発表ペーパー・要旨（国際会議）, 1880-6546, WOS:000322352200078 - Cerebellar LTD and regulation by TARPs
Toshihiro Nomura; Wataru Kakegawa; Shinji Matsuda; Kazuhisa Kohda; Michisuke Yuzaki
ELSEVIER IRELAND LTD, 出版日 2010年, NEUROSCIENCE RESEARCH, 68巻, 掲載ページ E342-E342, 英語, 研究発表ペーパー・要旨（国際会議）, 0168-0102, WOS:000208443702277 - New mechanisms regulating stability and dynamics of AMPA receptors
Michisuke Yuzaki; Shinji Matsuda
ELSEVIER IRELAND LTD, 出版日 2010年, NEUROSCIENCE RESEARCH, 68巻, 掲載ページ E7-E7, 英語, 研究発表ペーパー・要旨（国際会議）, 0168-0102, WOS:000208443700024 - New mechanisms regulating the number of AMPA receptors at synapses
Michisuke Yuzaki; Shinji Matsuda
ELSEVIER IRELAND LTD, 出版日 2009年, NEUROSCIENCE RESEARCH, 65巻, 掲載ページ S28-S28, 英語, 研究発表ペーパー・要旨（国際会議）, 0168-0102, WOS:000272421100188 - メンブレントラフィックと神経系 神経における極性輸送--ついに樹状突起特異的輸送の分子機構の一端が明らかに (メンブレントラフィックの奔流--分子から細胞,そして個体へ) -- (メンブレントラフィックと高次機能)
松田 信爾; 柚崎 通介
共立出版, 出版日 2008年12月, 蛋白質核酸酵素, 53巻, 16号, 掲載ページ 2214-2219, 0039-9450, 40016364877 - Disruption of AMPA receptor GluR2 clusters following long-term depression induction in cerebellar Purkinje neurons (vol 19, pg 2765, 2000)
S Matsuda; T Launey; S Mikawa; H Hirai
OXFORD UNIV PRESS, 出版日 2003年01月, EMBO JOURNAL, 22巻, 1号, 掲載ページ 174-174, 英語, その他, 0261-4189, WOS:000180435100017 - Phosphorylation of serine 880 in GluR2 by protein kinase C prevents its C-terminus binding with the glutamate receptor interacting protein, GR
S Matsuda; S Mikawa; H Hirai
BLACKWELL SCIENCE LTD, 出版日 2000年, EUROPEAN JOURNAL OF NEUROSCIENCE, 12巻, 掲載ページ 465-465, 英語, 研究発表ペーパー・要旨（国際会議）, 0953-816X, WOS:000088236602643
書籍等出版物
- CLINICAL NEUROSCIENCE 小脳学習説 Marr-Albus-Ito理論の50年
掛川渉; 松田信爾; 柚﨑通介
学術書, 日本語, 共著, 小脳学習における長期抑圧(LTD)の役割, 中外医学社, 出版日 2019年 - 脳科学辞典（アダプタータンパク質複合体）
松田信爾
事典・辞書, 日本語, 単著, アダプタータンパク質複合体, 出版日 2018年11月17日 - スタンフォード神経科学
Ligun Luo
教科書・概説・概論, 日本語, 共訳, 第3章, メディカルサイエンス・インターナショナル, 出版日 2017年08月08日 - 脳科学辞典（長期抑圧）
松田信爾
事典・辞書, 日本語, 単著, 長期抑圧, 日本神経科学学会, 出版日 2015年09月17日 - 蛋白質核酸酵素増刊 「メンブレントラフィックの奔流」
松田信爾; 柚崎通介
学術書, 日本語, 共著, 神経における極性輸送―ついに樹状突起特異的輸送の分子機構の一端が明らかに, 共立出版, 出版日 2008年12月
講演・口頭発表等
- リソソーム開口放出に着目した光遺伝学による長期可塑性の新規制御ツール
荒井 格; 掛川 渉; 今井 貴雄; 佐賀 一期; 吉住 玲; 角田 聡; 松田 恵子; 神取 秀樹; 松田 信爾; 柚﨑 通介
ポスター発表, 英語, 第46回日本神経科学大会
発表日 2023年08月03日
開催期間 2023年08月01日- 2023年08月04日 - Chemical LTD stimulation induced the drastic mitochondrial morphological changes in cultured hippocampal neurons
新 尚也; 松田 信爾
ポスター発表, 英語, 日本生理学会第100回記念大会, 査読付
発表日 2023年03月15日
開催期間 2023年03月14日- 2023年03月16日 - シナプス可塑性の光操作－記憶・学習を制御するオプトジェネティクス－
松田信爾
口頭発表（招待・特別）, 日本語, シナプス研究会, 招待, 生理学研究所, zoom開催, 国内会議
発表日 2020年12月03日 - シナプス可塑性の光操作－記憶・学習を制御するオプトジェネティクス－
松田信爾
口頭発表（招待・特別）, 日本語, 生理研シナプス研究会, 招待, 国内会議
発表日 2020年12月03日 - Lysosomal exocytosis controls AMPA receptor trafficking
松田信爾
シンポジウム・ワークショップパネル（指名）, 英語, 第43回日本神経科学大会, 国際会議
発表日 2020年07月 - MMP-9 activity is required for the NMDA induced endocytosis of AMPA receptor
Shinnosuke Kohara; Shinji Matsuda
ポスター発表, 英語, 9th FAOPS, Federation of the Asian and Oceanian Physiological Sosieties, Kobe convention center, 国際会議
発表日 2019年03月29日 - New optogenetical tool clarified that the cerebellar LTD was essential for motor learning
Shinji Matsuda
シンポジウム・ワークショップパネル（指名）, 英語, 9th FAOPS, 招待, Federation of the Asian and Oceanian Physiological societies, Kobe convention center, 国際会議
発表日 2019年03月29日 - Lysosomal exocytosis is essential for NMDA-induced AMPA receptor exocytosis and LTP in hippocampus
Shinji Matsuda; Itaru Arai; Micisuke Yuzaki
ポスター発表, 英語, Japan-UK Neuroscience Symposium 2019, 国際会議
発表日 2019年02月10日 - Optogenetical control of AMPA receptor endocytosis clarified that the cerebellar long term depression directly regulate motor learning
Shinji Matsuda
口頭発表（招待・特別）, 英語, UK-Japan Neuroscience Symposium, AMED, MRC, ロンドン, 国際会議
発表日 2018年03月05日 - Development of the optogennetical technique which control the endocytosis of AMPA receptor and the long-term depression
Shinji Matsuda
口頭発表（一般）, 英語, The 40th Annual Meeting of the Japan Neuroscience Society, 国際会議
発表日 2017年07月21日 - 小脳長期抑圧の制御技術開発と運動学習メカニズムの解析
松田信爾
口頭発表（一般）, 英語, 第94回日本生理学会
発表日 2017年03月30日 - Novel optogenetical technique clarified that the cerebellar LTD was essential for the motor learning
Shinji Matsuda
口頭発表（一般）, 英語, Wiring and Functional Principles of Neural Circuits, JST, University of California, San Diego,, 国際会議
発表日 2016年11月17日 - The interaction between stargazin and synaptotagmin-7 is controlled by both of phosphorylation and Ca2+ concentration.
髙橋亜紀代; 松田信爾
ポスター発表, 英語, The 39th Annual Meeting of the Japan Neuroscience Society, 日本神経科学学会, 横浜, 国際会議
発表日 2016年07月21日 - AMPA受容体－TARP複合体の細胞内輸送機構
松田信爾
口頭発表（一般）, 英語, 日本生理学会 日中合同シンポジウム, 招待, 日本生理学会, 札幌, 国際会議
発表日 2016年03月23日 - Understanding and controlling synaptic plasticity
Shinji Matsuda
口頭発表（一般）, 英語, 眼球運動を制御する小脳中枢機構の理解へのデータ駆動型アプローチ, 招待, 日本神経回路学会 電気通信大学 脳科学ライフサポート研究センター, 東京, 国際会議
発表日 2015年08月01日 - Molecular mechanism and controlling method for synaptic plasticity
Shinji Matsuda
シンポジウム・ワークショップパネル（公募）, 英語, 第38回日本神経科学大会, 日本神経科学会, 神戸コンベンションセンター, 国内会議
発表日 2015年07月30日 - シナプス可塑性を支えるAMPA受容体の細胞内輸送機構
松田信爾
口頭発表（一般）, 日本語, 第57回日本神経化学会大会, 国内会議
発表日 2014年09月29日 - Development of the technology for controlling synaptic plasticity
Shinji Matsuda; Michisuke Yuzaki
口頭発表（一般）, 英語, Neuro2013, 国際会議
発表日 2013年 - Stargazin regulates AMPA receptor trafficking from plasma membrane to early endosome and lysosome during long term depression.
Shinji Matsuda; Michisuke Yuzaki
口頭発表（一般）, 英語, The 90th Annual Meeting of the Physiological Society of Japan Korea-Japan-Chine Joint Symposium, Physiological Society of Japan, 国際会議
発表日 2013年 - AMPA受容体のエンドサイトーシス制御機構
松田信爾
口頭発表（招待・特別）, 日本語, 生理研研究会「シナプス可塑性の動作原理」, 招待, 生理研研究会, 国内会議
発表日 2012年06月14日 - New mechanisms regulating the number of AMPA receptors at synapses
Michisuke Yuzaki; Shinji Matsuda
口頭発表（一般）, 英語, Neuro2009, 国際会議
発表日 2009年 - AP-4によるAMPA受容体の極性輸送
松田信爾
口頭発表（招待・特別）, 日本語, 第１７回 海馬と高次脳機能学会 特別講演, 招待, 海馬と高次脳機能学会, 国内会議
発表日 2008年11月22日 - Mechanisms for polarized sorting of glutamate receptors to the somatodendritic domain of neurons
Shinji Matsuda; Eriko Miura; Wataru Kakegawa; Keiko Matsuda; Kazuhisa Kohda; Masahiko Watanabe; Michisuke Yuzaki
口頭発表（一般）, 英語, Neuro2008, に本神経科学学会, 国際会議
発表日 2008年 - Mechanisms for Polarized Sorting of Glutamate Receptors to the Somatodendritic Domain of Neurons
Shinji Matsuda; Keiko Matsuda; Eriko Miura; Masahiko Watanabe; Michisuke Yuzaki
口頭発表（一般）, 英語, The 84th Annual Meeting of the Physiological Society of Japan, Physiological Society of Japan, 国際会議
発表日 2007年
担当経験のある科目_授業
- 基盤理工学専攻基礎
The University of Electro-Communications - 基盤理工学専攻基礎
電気通信大学 - 化学生命工学実験第二
The University of Electro-Communications - 化学生命工学実験第二
電気通信大学 - UECパスポートセミナー
The University of Electro-Communications - UECパスポートセミナー
電気通信大学 - 生体情報システム論１
電気通信大学 - 生体情報システム論１
電気通信大学 - 先進理工学基礎
The University of Electro-Communications - 生体機能システム実験第二
電気通信大学 - 先進理工学基礎
電気通信大学 - 先進理工学基礎
電気通信大学 - 生体機能システム実験第二
The University of Electro-Communications - 生体機能システム実験第二
電気通信大学 - Biology
The University of Electro-Communications - Molecular Biology
The University of Electro-Communications - 分子生物学
電気通信大学 - neurosience
The University of Electro-Communications - 神経科学
電気通信大学 - 生物学
電気通信大学 - 生物学
電気通信大学 - Fundamentals of Information Biology
The University of Electro-Communications - 生体情報学基礎
電気通信大学
共同研究・競争的資金等の研究課題
- 光操作によるシナプス可塑性と記憶形成の因果関係の解明
柚﨑通介
科学技術振興機構, 戦略的な研究開発の推進 戦略的創造研究推進事業 CREST, 慶應義塾大学, 神経回路を構成するシナプスは、神経活動に応じて生涯に亘って可塑的に変化します。この現象は長期増強(LTP)や長期抑圧(LTD)として長年研究されてきました。しかし、特定のシナプスにおけるLTP/LTDが、個体レベルにおける記憶・学習と因果関係が本当にあるのか、という根源的な問いは未解決です。本研究では、急性かつ可逆的にシナプス可塑性を直接制御する光遺伝学的ツールを開発することによってこの問いに挑みます。
研究期間 2018年10月 - 2024年03月 - TARPのリン酸化によるAMPA受容体輸送の制御機構
松田信爾
日本学術振興会, 科学研究費女性事業, 研究代表者
研究期間 2017年04月01日 - 2020年03月31日 - 小脳をモデルとした糖鎖シグナルによる機能的・形態的シナプス可塑性制御
柚崎 通介; 松田 信爾; 松田 恵子
日本学術振興会, 科学研究費助成事業, 慶應義塾大学, 新学術領域研究(研究領域提案型), 中・長期に持続する記憶・学習は、シナプス後部におけるAMPA受容体の輸送によって担われ、より長期に持続する記憶はシナプスの形態変化が伴う。本研究では小脳神経回路をモデルとし、糖鎖科学と神経科学的アプローチを融合することによって、機能的・形態的シナプス可塑性を制御する分子機構の解明を目指した。まず、シナプス可塑性のモデルである長期抑圧や長期増強時には、AMPA受容体はライソゾーム様細胞内プールから出入りし、ライソゾームに存在する糖鎖分解酵素が大きな働きをすることが分かった。またプルキンエ細胞への子宮内電気穿孔法と時期特異的遺伝子操作法を確立し、樹状突起とシナプス形態形成過程の分子機構を解明した。, 23110009
研究期間 2011年04月01日 - 2016年03月31日 - 光による細胞内輸送とシナプス可塑性の制御
松田信爾
科学技術振興機構, 研究助成, 研究代表者
研究期間 2011年 - 2016年03月 - 細胞内輸送機構による神経機能の制御
松田信爾
文部科学省, 研究助成, 研究代表者
研究期間 2010年 - 2012年 - 神経細胞における樹状突起特異的輸送機構
松田信爾
文部科学省, 研究助成, 研究代表者
研究期間 2008年 - 2010年 - 神経細胞間で機能する新しい分泌性C1qファミリー分子群の解析
柚崎 通介; 松田 信爾; 飯島 崇利
日本学術振興会, 科学研究費助成事業, 慶應義塾大学, 特定領域研究, 補体Clqの球状ドメイン(gClq)を磯能ドメインとして持つClq/TNFスーパーファミリーが、シグナル分子や細胞外マトリックス分子として細胞外環境において非常に多彩な生理機能に関与することが近年注目を集めている。一方、Clq/TNFスーパーファミリーの中には脳内に主に発現するCblnファミリーとClq1ファミリーが存在するが、これらの分子の生理機能についてはほとんど分かっていない。私たちはこれまでにCblnファミリーのうちCbln1分子が、小脳顆粒細胞-プルキンエ細胞シナプスにおいて、シナプスの接着性と可塑性を制御することを発見した。Cbln1は海馬歯状回に線維を送る嗅内皮質にも発現し、他のメンバー分子Cbln2やCbln4も海馬や脳内の各部位に発現していることから、他の脳部位においてもCblnファミリーがシナプス形態調節分子として機能している可能性がある(Eur J Neurosci,in press)。今年度はClq1ファミリー分子群(Clql1-Clql4)の解析を進めた。Clql1は小脳登上線維の起始核である下オリーブ核に特異的に発現し、Clql2とClql3分子は海馬歯状回の顆粒細胞に発達期および成熟後も共発現する。これらの分子は何れもCblnファミリーと同様に同種・異種分子間にて多量体を形成して分泌されることを見いだした(Eur J Neurosci,2010)。これらのことからClqlファミリーもCblnファミリーと同様に、発達時や成熟後の脳においてシナプス機能に関与している可能性が示唆された。, 20057024
研究期間 2008年 - 2009年 - 神経細胞樹状突起へのAMPA受容体トランスポートソームの特異的輸送機構
柚崎 通介; 松田 信爾; 松田 恵子
日本学術振興会, 科学研究費助成事業, 慶應義塾大学, 特定領域研究, 神経細胞は、樹状突起と軸索という、機能的・構造的に大きく異なった領域をもった極性細胞である。神経細胞における速い興奮性情報伝達は、樹状突起上の細胞膜に存在するAMPA型グルタミン酸受容体によって担われている。しかしどのようにしてAMPA受容体が樹状突起へ選択的に輸送されるのかについては十分分かっていない。私たちはこれまでに、アダプタータンパク質AP-4が樹状突起への極性輸送を制御することを発見した。AMPA型グルタミン酸受容体はその関連タンパク質TARPに結合し、TARPをAP-4が認識することにより軸索方向への輸送を阻害する。一方、軸索に誤輸送されたAMPA受容体は細胞表面に輸送されず、軸索内部においてオートファジーにより分解されることが判明した。そこで今年度は、軸索内部においてどのようにオートファジー経路が制御されているのかを検討した。グルタミン酸受容体が過剰に活性化されると、細胞内に流入したNaイオンなどを排出するためにポンプ活性が亢進し、その結果ATP濃度の低下とAMPK-mTOR経路を介したオートファジーの活性化が起きることを見いだした(J Neurosci,2009)。したがって、神経変性モデルマウスであるラーチャーにおける軸索内部でのオートファジーは、神経細胞死の原因ではなく、結果でありむしろ細胞死を防ぐ方向に働いていると考えられた。一方、軸索で形成されたオートファゴゾームはダイニンによって細胞体に逆行性輸送されることを見いだした(Autophagy,2009)。, 20056028
研究期間 2008年 - 2009年 - 神経細胞における極性輸送の分子機構
松田信爾
文部科学省, 研究助成, 研究代表者
研究期間 2006年 - 2008年 - 神経細胞における極性輸送を支える新しいトラフィック制御機構
柚崎 通介; 松田 信爾; 松田 恵子
日本学術振興会, 科学研究費助成事業, 慶應義塾大学, 特定領域研究, 神経細胞は、樹状突起と軸索という、機能的・構造的に大きく異なった領域をもった極性細胞である。哺乳類の神経細胞における情報伝達は、軸索末端から放出されるグルクミン酸が、樹状突起上の細胞膜に存在するグルタミン酸受容体に結合することにより成立する。したがって、神経細胞の極性とその維持は、神経細胞の情報伝達に必須である。しかし神経細胞において極性にしたがって膜タンパク質が樹状突起や軸索へ選択的に輸送される分子機構については、ほとんど分かっていなかった。 私たちは、アダプタータンパク質AP-4が樹状突起への極性輸送を制御することを世界に先駆けて明らかにした(Neuron 57:730, 2008)。 AMPA型グルタミン酸受容体はその関連タンパク質TARPに結合し、TARPをAP-4が認識することにより軸索方向への輸送を阻害する。したがってAP-4ノックアウトマウスにおいてはTARPとAMPA型グルタミン酸受容体は軸索方向に誤輸送された。同様に、リポタンパク質受容体LDLRはAP-4ノックアウトマウスにおいて軸索に誤輸送された。一方、NMDA型グルタミン酸受容体、代謝型グルタミン酸受容体、トランスフェリン受容体TfRは、AP-4ノックアウトマウスにおいても樹状突起に選択的に輸送されたことから、AP-4に依存しない極性輸送系が存在することが明らかとなった。面白いことに、軸索に誤輸送された膜タンパク質は細胞表面に輸送されず、軸索内部においてオートファジーにより分解されることが判明し、AP-4がオートファジー経路にも関わっている可能性が示唆され、, 18050034
研究期間 2006年 - 2007年 - 成熟脳におけるシナプス維持と記憶の形成-δ2グルタミン酸受容体を介する新しい機構
柚崎 通介; 飯島 崇利; 松田 信爾
日本学術振興会, 科学研究費助成事業, 慶應義塾大学, 基盤研究(A), 神経伝達物質であるグルタミン酸は、シナプス形成や記憶学習、さらには神経細胞死などの多彩な機能に関与する。δ2受容体は、小脳平行線維がプルキンエ細胞と形成するシナプス後膜に特異的に発現しており、このシナプスにおける可塑性過程である長期抑圧現象(LTD)を制御する。しかし、δ2受容体のリガンドが不明であるために、δ2受容体を介する信号伝達経路の解明は遅れている。これまでの研究により、グルタミン酸受容体ファミリーにおいて進化的にも完全に保存されているリガンド結合コンセンサス部位に変異を加えても、δ2受容体の小脳における機能(協調運動・平行線維シナプス形成・シナプス可塑性)が保たれることを見出した(EMBO Rep,2005)。一方、δ2受容体には、他のグルタミン酸受容体においてチャネルを構成するアミノ酸残基が高度に保存されているが、チャネル機能を持っているかどうかは明らかでなかった。そこで、チャネルポア部分のアミノ酸残基を変異させることにより、チャネルのCa^<2+>イオン透過性やイオン透過性全般を消失した変異δ2受容体を発現するトランスジェニックマウスやウイルスベクターを作成した。驚くべき事に、これらの変異δ2受容体は正常δ2受容体と全く同じ機能を有することから、δ2受容体は、「イオンチャネル型グルタミン酸受容体」に属するものの、チャネルとして働かないことが明らかになった(J Physiol,2007)。一方、δ2受容体の細胞内ドメインの最C末端部分を欠損させるとその機能を失うことから、δ2受容体はC末端部分に結合するタンパク質を介してシグナル伝達を行う「代謝型活性」を持つことが示唆された(Eur J Neurosci,in press)。, 16200024
研究期間 2004年 - 2006年 - 小脳における記憶・学習の新しい分子機構―δ2グルタミン酸受容体の役割
松田信爾
文部科学省, 研究助成, 研究代表者
研究期間 2004年 - 2006年 - グルタミン酸受容体の構造解析のための新しい精製法の開発-安定多量体ペプチドの応用
柚崎 通介; 松田 信爾
日本学術振興会, 科学研究費助成事業, 慶應義塾大学, 萌芽研究, 哺乳類の中枢神経系の速い神経伝達はグルタミン酸受容体チャネル(GluR)により担われる。更にGluRはシナプス形成や記憶学習、さらには神経細胞死などの多彩な神経機能に関与する。従って、生理的・病的状態におけるシナプス伝達機構を理解するためには、GluRの構造を解明することが非常に重要な課題となっている。しかし膜蛋白質、とりわけGluRのような多量体構造を取る蛋白質の精製は容易ではない。本研究では、GluRのN末端部に、非常に安定した多量体構造を取る人工ペプチドを付加するという新しいアイデアにより、GluRを構造的に安定させて構造解析に適したGluRの精製法を開発することを目的とした。 33アミノ酸からなるGCN4ペプチドは、ロイシンジッパー構造を持ち、高温でも非常に安定な4量体を形成することが知られている。また、ロイシンジッパー部位の数ヶ所のアミノ酸を変異させると、今度は非常に安定な2量体、3量体を形成する。そこで、GluRのN末端部にこれらの人工ペプチドを付加し、その構造と機能について検討した。GluR1のN末端部にGCN4ペプチドを導入すると、その多量体構造に応じて、GluR1チャネル複合体におけるGluR1サブユニットの数が正確にコントロールできた。いずれのGluR1変異体も、細胞表面に正常に輸送されるが、4量体型GCN4をN末端部に導入した時にのみ、正常のチャネル機能が観察されることを電気生理学的に確認した。これらのことから、GluR1は4量体構造をとること、N末端側に4量体化ペプチドを導入したGluR1は正常機能を持つことが確認できた(J Biol Chem,2005)。この結果を活かして、GluRの精製法をさらに検討中である。, 16657047
研究期間 2004年 - 2005年 - 神経細胞における受容体の選択的輸送とシナプス可塑性を支えるトラフィック制御機構
柚崎 通介; 幸田 和久; 松田 恵子; 松田 信爾
日本学術振興会, 科学研究費助成事業, 慶應義塾大学, 特定領域研究, 神経細胞は高度に発達した極性を持った細胞であり、軸索と樹状突起にはそれぞれ特異的な膜蛋白質が発現している。また、記憶・学習の基礎過程である長期抑圧現象(LTD)は、シナプス後膜に存在するグルタミン酸受容体の選択的エンドサイトーシスにより起きることが、近年明らかになった。このように神経細胞機能の理解には、膜蛋白質のトラフィック機構の解明が重要な鍵を握る。私たちは小脳プルキンエ細胞樹状突起に特異的に発現し、LTD発現と小脳での学習機能を制御するδ2型グルタミン酸受容体をモデルとして、トラフィック機構を解明することにより、神経細胞における高次機能の発現機構を明らかにすることを目的とした。 これまでに、δ2受容体の細胞内C末端部分に、小胞体からトランスゴルジ網に輸送させるA領域(Eur.J.Neurosci.,'04)、シナプス後膜からのエンドサイトーシスを妨げるE領域(投稿中)、樹状突起への極性輸送を制御するF領域がそれぞれ独立して存在することを明らかにした。一方、細胞外のN末端領域は、輸送には直接関係しないものの、安定多量体形成に必要であり、その障害によりδ2受容体の小胞体への貯留が起きることを見出した(J.Biol.Chem,'05)。さらに、F領域についてさらに解析を進め、この領域に結合することにより樹状突起への極性輸送を制御する蛋白質を同定した。この蛋白質のノックダウンにより、δ2受容体のみならずAMPA型グルタミン酸受容体の樹状突起への選択的輸送が障害されることから、この蛋白質は神経細胞における膜蛋白質の極性輸送を制御するマスター分子の一つであると考えられる(投稿準備中)。, 16044241
研究期間 2004年 - 2005年 - 小脳における記憶・学習の新しい分子機構―δ２グルタミン酸受容体の役割
慶應医師会医学研究助成金
研究期間 2003年 - 2004年03月31日 - AMPA型グルタミン酸受容体のリン酸化による神経細胞の感度調節機構
松田信爾
文部科学省, 研究助成, 研究代表者
研究期間 2000年 - 2000年