理学博士, 大阪大学

Ph.D, Osaka University

synaptic plasticity, long term depression (LTD), long term potentiation (LTP), endocytosis, exocytosis

Life sciences, Neuroscience - general

01 Nov. 2014
University of electro-communication, Graduate School of Informatics and Engineering, Associate Professor

01 Apr. 2009 - 31 Oct. 2014
Keio University, Department of Physiology School of Medicine, Assistant Professor

01 Apr. 2007 - 31 Mar. 2009
Keio University, Department of Physiology School of Medicine, Instructor

01 Oct. 2003 - 31 Mar. 2009
Keio University, Department of Physiology School of Medicine, Instructor

01 Oct. 2000 - 30 Sep. 2003
St. Jude Children's Research Hospital, Developmental Neurobiology, Postdoctral Fellow

01 Apr. 1998 - 30 Sep. 2000
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

01 Jul. 1995 - 31 Mar. 1998
日本学術振興会, 特別研究員ＤＣ１

01 Apr. 1995 - 25 Mar. 1998
大阪大学大学院, 理学研究科, 生物化学専攻

01 Apr. 1993 - 31 Mar. 1995
大阪大学大学院, 理学研究科, 生物化学専攻

01 Apr. 1989 - 31 Mar. 1993
大阪大学, 理学部, 生物学科

Mar. 1989
大阪府立大手前高校, 普通科

Mar. 2021
評議員, 日本小脳学会, Society

2019
プログラム委員, 日本神経科学学会, Society

Apr. 2018
評議員, 日本生理学会

Aug. 2018
Elsevier
Outstanding Reviewer
Publisher

2008
に本神経科学学会
日本神経科学学会奨励賞
Japan society

SARM1 is essential for NMDA receptor-dependent endocytosis of AMPA receptors in hippocampal neurons.
Misaki Morishita; Shinji Matsuda
Last, Neuroscience research, 28 Sep. 2024, Peer-reviwed, True, Long-term depression (LTD) is a form of synaptic plasticity thought to be the cellular basis of experience-dependent learning and memory. LTD is caused by an activity-dependent decrease in cell surface α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA)-type glutamate receptors (AMPA receptors) at the postsynaptic sites. However, the mechanism through which AMPA receptors are removed from the cell surface via neuronal activity is not fully understood. In this study, we showed that small interfering RNA (siRNA)-mediated knockdown of sterile alpha and toll/interleukin receptor motif containing 1 (SARM1) in cultured hippocampal neurons prevented the N-methyl-d-aspartate (NMDA)-induced reduction in cell surface AMPA receptors. However, the control RNA did not affect NMDA-mediated AMPA receptor trafficking. Overexpression of the siRNA-resistant form of SARM1 in SARM1-knocked-down neurons restored AMPA receptor trafficking. However, overexpression of SARM1, which lacks the mitochondrial transport signal, in the SARM1-knocked-down neurons did not restore NMDA-dependent AMPA receptor endocytosis. Moreover, the inhibition of the NADase activity of SARM1 blocked the NMDA-induced reduction of cell surface AMPA receptors. These results suggest that both the mitochondrial localization and NADase activity of SARM1 are essential for NMDA receptor-dependent AMPA receptor internalization in the hippocampal neurons.
Scientific journal, English

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
Last, Journal of Biological Chemistry, Elsevier BV, 300, 3, 105744-105744, Mar. 2024, Peer-reviwed
Scientific journal, English

Subunit-dependent and subunit-independent rules of AMPA receptor trafficking during chemical long-term depression in hippocampal neurons
Shinji Matsuda; Michisuke Yuzaki
Lead, Journal of Biological Chemistry, Cold spring harbor laboratory, 297, 2, 100949, 10 Jul. 2021, Peer-reviwed
Scientific journal, English

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, 4305, 1-15, 13 Mar. 2019, Peer-reviwed
Scientific journal, English

PhotonSABER: new tool shedding light on endocytosis and learning mechanisms in vivo.
Shinji Matsuda; Wataru Kakegawa; Michisuke Yuzaki
Lead, Communicative & integrative biology, Taylor & Francis, 12, 1, 34-37, 2019, Peer-reviwed, Invited, True, 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.
Scientific journal, English

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, 10 Sep. 2018, Invited
Scientific journal, Japanese

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
Last, Neuron, Cell Press, 99, 5, 985-998, 05 Sep. 2018, Peer-reviwed, True, 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.
Scientific journal, English

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, 14850, Apr. 2017, Peer-reviwed, 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.
Scientific journal, English

AP-4
Shinji Matsuda; MIchisuke Yuzaki
Encyclopedia of Signaling Molecules 2nd edition, Springer New York, 1-6, 2016, Invited
Scientific journal, English

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, 2, 422-431, Jan. 2014, Peer-reviwed, 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.
Scientific journal, English

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
Lead, NATURE COMMUNICATIONS, NATURE PUBLISHING GROUP, 4, 2759, Nov. 2013, Peer-reviwed, 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.
Scientific journal, English

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, 10, E948-E957, Mar. 2013, Peer-reviwed, 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.
Scientific journal, English

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, 1, 135-148, Jan. 2012, Peer-reviwed, 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.
Scientific journal, English

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, 5, 603-U93, May 2011, Peer-reviwed, 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.
Scientific journal, English

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, 402-410, 2011, Peer-reviwed
Scientific journal, English

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, 2-3, 694-694, Aug. 2010
English

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, Suppl., 67-67, Mar. 2009
Japanese

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, 4, 707-717, Feb. 2009, Peer-reviwed, 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.
Scientific journal, English

Polarized sorting of AMPA receptors to the somatodendritic domain is regulated by adaptor protein AP-4.
Shinji Matsuda; Mishisuke Yuzaki
Neuroscience Research, 65, 1-5, 2009, Peer-reviwed
Scientific journal, English

AP-4: Autophagy-four mislocalized proteins in axons
Shinji Matsuda; Michisuke Yuzaki
Autophagy, Taylor and Francis Inc., 4, 6, 815-816, 16 Aug. 2008, Peer-reviwed, Invited, 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.
Scientific journal, English

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, 5, 730-745, Mar. 2008, Peer-reviwed, 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.
Scientific journal, English

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, S21, 2008, Peer-reviwed

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, 5, 1357-1362, Mar. 2007, Peer-reviwed, 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.
Scientific journal, English

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, 35, 25577-25587, Sep. 2006, Peer-reviwed, 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.
Scientific journal, English

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, 25, 17501-17509, Jun. 2006, Peer-reviwed, 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.
Scientific journal, English

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, 20, 20021-20029, May 2005, Peer-reviwed, 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.
Scientific journal, English

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, 1, 90-95, Jan. 2005, Peer-reviwed, 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.
Scientific journal, English

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, 7, 1683-1690, Apr. 2004, Peer-reviwed, 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.
Scientific journal, English

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, 8, 1581-1590, Apr. 2003, Peer-reviwed, 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.
Scientific journal, English

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, 1, 27-37, Jan. 2003, Peer-reviwed, 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.
Scientific journal, English

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, 8, 1507-1516, Oct. 2002, Peer-reviwed, 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.
Scientific journal, English

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, 43-52, 2002, Peer-reviwed
Scientific journal, English

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, 12, 2765-2774, Jun. 2000, Peer-reviwed, 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.
Scientific journal, English

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, 4, 1765-1768, Oct. 1999, Peer-reviwed, 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.
Scientific journal, English

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, 4, 281-287, Sep. 1999, Peer-reviwed, 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.
Scientific journal, English

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, 3, 157-163, Aug. 1999, Peer-reviwed, 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.
Scientific journal, English

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, 4, 1310-1315, Jan. 1999, Peer-reviwed, 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.
Scientific journal, English

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, 44-53, 1999, Peer-reviwed, 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.
Scientific journal, English

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, 32, 20223-20227, Aug. 1998, Peer-reviwed, 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.
Scientific journal, English

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, 3, 159-164, Mar. 1998, Peer-reviwed, 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.
Scientific journal, English

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, 1, 173-177, May 1997, Peer-reviwed, 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.
Scientific journal, English

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, 35, 21359-21364, Aug. 1996, Peer-reviwed, 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.
Scientific journal, English

Chemical LTD stimulation induced the drastic mitochondrial morphological changes in cultured hippocampal neurons
Naoya Atarashi; Shinji Matsuda
Last, Mar. 2023, The Journal of Physiological Sciences, 73, 11, 130-130, English, Summary national conference

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
Dec. 2022, The Journal of Physiological Sciences, 72, 33, 151-151, English, Summary national conference

【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, Aug. 2019, Clinical Neuroscience, 37, 8, 919-924, Japanese, 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, Nov. 2014, GLYCOBIOLOGY, 24, 11, 1201-1201, English, Summary international conference, 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, 2013, JOURNAL OF PHYSIOLOGICAL SCIENCES, 63, S28-S28, English, Summary international conference, 1880-6546, WOS:000322352200045

How does the delta 2 glutamate receptor regulate cerebellar LTD?
Kazuhisa Kohda; Wataru Kakegawa; Shinji Matsuda
SPRINGER JAPAN KK, 2013, JOURNAL OF PHYSIOLOGICAL SCIENCES, 63, S38-S38, English, Summary international conference, 1880-6546, WOS:000322352200078

Cerebellar LTD and regulation by TARPs
Toshihiro Nomura; Wataru Kakegawa; Shinji Matsuda; Kazuhisa Kohda; Michisuke Yuzaki
ELSEVIER IRELAND LTD, 2010, NEUROSCIENCE RESEARCH, 68, E342-E342, English, Summary international conference, 0168-0102, WOS:000208443702277

New mechanisms regulating stability and dynamics of AMPA receptors
Michisuke Yuzaki; Shinji Matsuda
ELSEVIER IRELAND LTD, 2010, NEUROSCIENCE RESEARCH, 68, E7-E7, English, Summary international conference, 0168-0102, WOS:000208443700024

New mechanisms regulating the number of AMPA receptors at synapses
Michisuke Yuzaki; Shinji Matsuda
ELSEVIER IRELAND LTD, 2009, NEUROSCIENCE RESEARCH, 65, S28-S28, English, Summary international conference, 0168-0102, WOS:000272421100188

How polarized sorting to neuronal dendrites is achieved: a newly discovered AP4-based system
松田 信爾; 柚崎 通介
共立出版, Dec. 2008, Protein, nucleic acid and enzyme, 53, 16, 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, Jan. 2003, EMBO JOURNAL, 22, 1, 174-174, English, Others, 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, 2000, EUROPEAN JOURNAL OF NEUROSCIENCE, 12, 465-465, English, Summary international conference, 0953-816X, WOS:000088236602643

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
Scholarly book, Japanese, Joint work, 小脳学習における長期抑圧(LTD)の役割, 中外医学社, 2019

脳科学辞典（アダプタータンパク質複合体）
松田信爾
Dictionary or encycropedia, Japanese, Single work, アダプタータンパク質複合体, 17 Nov. 2018

Principles of Neurobiology
Ligun Luo
Textbook, Japanese, Joint translation, 第3章, メディカルサイエンス・インターナショナル, 08 Aug. 2017

脳科学辞典（長期抑圧）
松田信爾
Dictionary or encycropedia, Japanese, Single work, 長期抑圧, 日本神経科学学会, 17 Sep. 2015

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
Scholarly book, Japanese, Joint work, 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, Dec. 2008

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
Poster presentation, English, 第46回日本神経科学大会
03 Aug. 2023
01 Aug. 2023- 04 Aug. 2023

Chemical LTD stimulation induced the drastic mitochondrial morphological changes in cultured hippocampal neurons
新 尚也; 松田 信爾
Poster presentation, English, 日本生理学会第100回記念大会, Peer-reviewed
15 Mar. 2023
14 Mar. 2023- 16 Mar. 2023

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
Invited oral presentation, Japanese, シナプス研究会, Invited, 生理学研究所, zoom開催, Domestic conference
03 Dec. 2020

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
Invited oral presentation, Japanese, 生理研シナプス研究会, Invited, Domestic conference
03 Dec. 2020

Lysosomal exocytosis controls AMPA receptor trafficking
Shinji Matsuda
Nominated symposium, English, The 43rd Annual Meeting of the Japan Neuroscience Society, International conference
Jul. 2020

MMP-9 activity is required for the NMDA induced endocytosis of AMPA receptor
Shinnosuke Kohara; Shinji Matsuda
Poster presentation, English, 9th FAOPS, Federation of the Asian and Oceanian Physiological Sosieties, Kobe convention center, International conference
29 Mar. 2019

New optogenetical tool clarified that the cerebellar LTD was essential for motor learning
Shinji Matsuda
Nominated symposium, English, 9th FAOPS, Invited, Federation of the Asian and Oceanian Physiological societies, Kobe convention center, International conference
29 Mar. 2019

Lysosomal exocytosis is essential for NMDA-induced AMPA receptor exocytosis and LTP in hippocampus
Shinji Matsuda; Itaru Arai; Micisuke Yuzaki
Poster presentation, English, Japan-UK Neuroscience Symposium 2019, International conference
10 Feb. 2019

Optogenetical control of AMPA receptor endocytosis clarified that the cerebellar long term depression directly regulate motor learning
Shinji Matsuda
Invited oral presentation, English, UK-Japan Neuroscience Symposium, AMED, MRC, London, International conference
05 Mar. 2018

Development of the optogennetical technique which control the endocytosis of AMPA receptor and the long-term depression
Shinji Matsuda
Oral presentation, English, The 40th Annual Meeting of the Japan Neuroscience Society, International conference
21 Jul. 2017

Analysis of the relationship between cerebellar LTD and motor learning by novel optogenetical tool
Shinji Matsuda
Oral presentation, English, The 94th Annual Meeting of the Physiological Society of Japan
30 Mar. 2017

Novel optogenetical technique clarified that the cerebellar LTD was essential for the motor learning
Shinji Matsuda
Oral presentation, English, Wiring and Functional Principles of Neural Circuits, JST, University of California, San Diego,, International conference
17 Nov. 2016

The interaction between stargazin and synaptotagmin-7 is controlled by both of phosphorylation and Ca2+ concentration.
Akiyo Takahashi; Shinji Matsuda
Poster presentation, English, The 39th Annual Meeting of the Japan Neuroscience Society, 日本神経科学学会, 横浜, International conference
21 Jul. 2016

Intracellular trafficking of AMPA receptor-TARP complex
Shinji Matsuda
Oral presentation, English, Annual Meeting of the Physiological society of Japan Japan-China Joint Symposium, Invited, Physiological Society of Japan, 札幌, International conference
23 Mar. 2016

Understanding and controlling synaptic plasticity
Shinji Matsuda
Oral presentation, English, Data-driven approach for understanding cerebellar mechanisms on eye movement control, Invited, 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, International conference
01 Aug. 2015

Molecular mechanism and controlling method for synaptic plasticity
Shinji Matsuda
Public symposium, English, Neuro2015, Janap neuroscience society, Kobe Convention Center, Domestic conference
30 Jul. 2015

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
Oral presentation, Japanese, 第57回日本神経化学会大会, Domestic conference
29 Sep. 2014

Development of the technology for controlling synaptic plasticity
Shinji Matsuda; Michisuke Yuzaki
Oral presentation, English, Neuro2013, International conference
2013

Stargazin regulates AMPA receptor trafficking from plasma membrane to early endosome and lysosome during long term depression.
Shinji Matsuda; Michisuke Yuzaki
Oral presentation, English, The 90th Annual Meeting of the Physiological Society of Japan Korea-Japan-Chine Joint Symposium, Physiological Society of Japan, International conference
2013

AMPA受容体のエンドサイトーシス制御機構
松田信爾
Invited oral presentation, Japanese, 生理研研究会「シナプス可塑性の動作原理」, Invited, 生理研研究会, Domestic conference
14 Jun. 2012

New mechanisms regulating the number of AMPA receptors at synapses
Michisuke Yuzaki; Shinji Matsuda
Oral presentation, English, Neuro2009, International conference
2009

AP-4によるAMPA受容体の極性輸送
松田信爾
Invited oral presentation, Japanese, 第１７回 海馬と高次脳機能学会 特別講演, Invited, 海馬と高次脳機能学会, Domestic conference
22 Nov. 2008

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
Oral presentation, English, Neuro2008, に本神経科学学会, International conference
2008

Mechanisms for Polarized Sorting of Glutamate Receptors to the Somatodendritic Domain of Neurons
Shinji Matsuda; Keiko Matsuda; Eriko Miura; Masahiko Watanabe; Michisuke Yuzaki
Oral presentation, English, The 84th Annual Meeting of the Physiological Society of Japan, Physiological Society of Japan, International conference
2007

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
The University of Electro-Communications

生体情報システム論１
The University of Electro-Communications

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
The University of Electro-Communications

先進理工学基礎
The University of Electro-Communications

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
The University of Electro-Communications

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

日本小脳学会

Society for Neuroscience

The Japan Neuroscience Society

Physiological society of Japan

Functions of toll-like receptors in synaptic plasticity
松田 信爾
Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research, The University of Electro-Communications, Grant-in-Aid for Scientific Research (C), 24K09694
Apr. 2024 - Mar. 2027

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
Michisuke Yuzaki
JST, 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。
Oct. 2018 - Mar. 2024

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, Principal investigator
01 Apr. 2017 - 31 Mar. 2020

Sugar chain-based signals regulating morphological and functional synaptic plasticity in the cerebellum
Yuzaki Michisuke; MATSUDA SHINJI; MATSUDA KEIKO
Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research, Keio University, Grant-in-Aid for Scientific Research on Innovative Areas (Research in a proposed research area), It is believed that long-term memory is initially stored as changes in the number of AMPA receptors at the postsynaptic sites and eventually converted into morphological changes at synapses for longer-term memory. However, molecular mechanisms underlying these processes have not been fully understood yet. Dendrites are surrounded by extracellular matrix (ECM) proteins, which are enriched with various sugar chains. Thus, AMPA receptor trafficking and morphological changes must be accompanied with changes of the surrounding ECM. In this study, we aimed to clarify how functional and morphological synaptic plasticity is regulated by the ECM in the cerebellum. We found that changes in neuronal activity induced AMPA receptor trafficking to/from lysosome-like organelle, which contained various kinds of glycosydase. Using a temporally-controlled gene knockout method, we also clarified how a single transcription factor regulated morphogenesis of spines and dendrites of Purkinje cells in vivo., 23110009
01 Apr. 2011 - 31 Mar. 2016

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, Principal investigator
2011 - Mar. 2016

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, Principal investigator
2010 - 2012

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, Principal investigator
2008 - 2010

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
2008 - 2009

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
2008 - 2009

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, Principal investigator
2006 - 2008

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
2006 - 2007

Maintenance of synapses and formation of memory in the adult brain : new mechanisms regulated by the delta2 glutamate receptor
YUZAKI Michisuke; IIJIMA Takatoshi
Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research, KEIO UNIVERSITY, Grant-in-Aid for Scientific Research (A), The ionotropic glutamate receptors play important roles in not only neurotransmission, but also synaptic plasticity and neurotoxicity in the mammalian brain. They are subdivided into four subfamilies: alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptors, kainate receptors, N-methyl-D-aspartate receptors, and delta glutamate receptors. The delta2 glutamate receptor (GluRd2), which is predominantly expressed in Purkinje cells, plays a crucial role in cerebellar functions : mutant mice with a disrupted GluRd2 gene (delta2-null mice) display impaired synapse formation and abrogated long-term depression (LTD), a form of synaptic underlying motor learning. Despite its importance, the mechanisms by which delta2 regulates cerebellar functions have remained elusive. Several fundamental questions, such as whether GluRd2 is activated by glutamate and whether GluRd2 forms ion channels, have remained elusive largely because there were no specific pharmacologic tools to manipulate the function of GluRd2. To circumvent the problem caused by the lack of pharmacologic tools, we employed a "transgenic rescue" approach by exploiting the fact that the several important regions, such as the putative ligand-binding and the channel pore domains, of GluRd2 shares considerable similarity with those of AMPA receptors. We introduced a mutant GluRd2 transgene, in which the conserved residues in the putative ligand-binding or channel pore domains were disrupted, into GluRd2-null Purkinje cells by either virus vectors or transgenic mice. Surprisingly, the mutant GluRd2 transgenes were as effective as the wild-type GluRd2 in rescuing the GluRd2-null mice. Thus, we propose that GluRd2 may not be activated by glutamate and may not function as an ion channel although it belongs to the "ionotropic glutamate receptor" family. In contrast, abrogated LTD was not restored in GluRd2-null Purkinje cells by the transduction of a mutant GluRd2 transgene lacking the C-terminal domain. Therefore, GluRd2 may function by modulating an intracellular signalling pathway through its C-terminal intracellular domain, to which several adaptor proteins bind., 16200024
2004 - 2006

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, Principal investigator
2004 - 2006

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
2004 - 2005

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
2004 - 2005

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
2003 - 31 Mar. 2004

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, Principal investigator
2000 - 2000

夏休みは電通大でラボ体験2023
Lecturer, 匠ガールプロジェクト, Research advise
15 Jul. 2023 - 15 Jul. 2023

スプリングスクール
Lecturer, 電気通信大学 脳・医工学研究センター
27 Mar. 2023 - 27 Mar. 2023

第47回日本神経科学大会プログラム委員
Competition etc, Planning etc, 日本神経科学学会, 01 Jun. 2023 - 27 Jul. 2024

博士前期課程入試面接
Review, 18 Aug. 2023 - 18 Aug. 2023

学域一般入試採点(後期試験）
Others, Others, 12 Mar. 2023 - 13 Mar. 2023

先端工学基礎課程面接試験
Others, 25 Nov. 2022 - 25 Nov. 2022