Study on molecular mechanism of cerebellar circuit formation

• Project/Area Number: 25290021
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Partial Multi-year Fund
• Section: 一般
• Research Field: Neurochemistry/Neuropharmacology
• Research Institution: Shinshu University
• Principal Investigator: UEMURA Takeshi 信州大学, 学術研究院医学系, 准教授 (00372368)
• Co-Investigator(Renkei-kenkyūsha): YOSHIDA Tomoyuki 富山大学, 医学薬学研究院, 准教授 (90372367)
• Project Period (FY): 2013-04-01 – 2017-03-31
• Project Status: Completed (Fiscal Year 2016)
• Budget Amount: ¥15,340,000 (Direct Cost: ¥11,800,000、Indirect Cost: ¥3,540,000); Fiscal Year 2015: ¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000); Fiscal Year 2014: ¥6,110,000 (Direct Cost: ¥4,700,000、Indirect Cost: ¥1,410,000); Fiscal Year 2013: ¥5,070,000 (Direct Cost: ¥3,900,000、Indirect Cost: ¥1,170,000)
• Keywords: 脳・神経 / 小脳 / 神経回路網構築 / 神経回路網形成

Outline of Final Research Achievements

Synapses are formed and maintained by trans-synaptic interaction between pre- and postsynaptic cell adhesion molecules. In the cerebellum, the trans-synaptic interaction of postsynaptic glutamate receptor GluRδ2 and presynaptic neurexins (Nrxns) through cerebellin precursor protein 1 (Cbln1) mediates parallel fiber (PF)-Purkinje cell (PC) synapse formation. In mammals, Nrxns are encoded by three genes (Nrxn1, Nrxn2, and Nrxn3). In this study, we generated cerebellar granule cell (GC)-specific Nrxn1, Nrxn2, and Nrxn3 triple KO mice. The cerebellar GCs-specific Nrxns triple KO mice showed severe ataxia and reduced cerebellar size. Histological analysis revealed that the number of cerebellar GCs was dramatically decreased in the cerebellar GCs-specific Nrxns triple KO mice. Electron microscopic analysis showed that PF-PC synapses were disappeared in the GCs-specific Nrxns triple KO mice. These results suggest that Nrxns are essential for cerebellar circuit formation.

Research Products

• [Journal Article] In situ screening for postsynaptic cell adhesion molecules during synapse formation (2017)
  • Author(s): Uemura T, Maeda A, Yasumura M, Shimada T, Fukata Y, Fukata M, Yoshida T
  • Journal Title: J Biochem Volume: 印刷中 Pages: 295-302
  • DOI: 10.1093/jb/mvx030
  • Peer Reviewed
• [Journal Article] Fluorescent protein tagging of endogenous protein in brain neurons using CRISPR/Cas9-mediated knock-in and in utero electroporation techniques (2016)
  • Author(s): Uemura T, Mori T, Kurihara T, Kawase S, Koike R, Satoga M, Cao X, Li X, Yanagawa T, Sakurai T, Shindo T, Tabuchi K.
  • Journal Title: Sci.Rep. Volume: 6 Pages: 35861-35861
  • DOI: 10.1038/srep35861
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Mechanisms of splicing-dependent trans-synaptic adhesion by PTPδ–IL1RAPL1/IL-1RAcP for synaptic differentiation. (2015)
  • Author(s): Yamagata A., Yoshida, T., Sato, Y., Goto-Ito, S., Uemura, T., Maeda, A., Shiroshima, T., Iwasawa-Okamoto, S., Mori, H., Mishina, M., Fukai , S.
  • Journal Title: Nat Commun. Volume: 6 Pages: 6926-6926
  • DOI: 10.1038/ncomms7926
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Expansion of stochastic expression repertoire by tandem duplication in mouse Protocadherin-α cluster. (2014)
  • Author(s): Misato Yasumura, Tomoyuki Yoshida, Maya Yamazaki, Manabu Abe, Rie Natsume, Kouta Kanno, Takeshi Uemura, Keizo Takao, Kenji Sakimura, Takefumi Kikusui, Tsuyoshi Miyakawa, Masayoshi Mishina
  • Journal Title: Sci Rep. Volume: 4: 6613 Pages: 6613-6613
  • DOI: 10.1038/srep06613
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Striatal dopamine D1 receptor is essential for contextual fear conditioning. (2014)
  • Author(s): Masaru Ikegami, Takeshi Uemura, Ayumi Kishioka, Kenji Sakimura and Masayoshi Mishina
  • Journal Title: Sci Rep. Volume: 4 Pages: 3976-3976
  • DOI: 10.1038/srep03976
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Consolidation of auditory fear memories formed by weak unconditioned stimuli requires NMDA receptor activation and de novo protein synthesis in the striatum (2013)
  • Author(s): Kishioka A, Uemura T, Fukushima F, Mishina M
  • Journal Title: Mol. Brain Volume: 6 Pages: 17-17
  • DOI: 10.1186/1756-6606-6-1
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Molecular Mechanism of Synapse Formation by Synaptic Organizer (2013)
  • Author(s): 植村 健
  • Journal Title: 生物物理 Volume: 53 Issue: 2 Pages: 090-093
  • DOI: 10.2142/biophys.53.090
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Presentation] RISPR/Cas9-mediated gene knock-in in single neurons in vivo (2016)
  • Author(s): 植村健, 森琢磨, 栗原大河, 里賀みちる, 田渕克彦
  • Organizer: 第39回日本神経科学大会
  • Place of Presentation: 横浜市、パシフィコ横浜
  • Year and Date: 2016-07-21
• [Presentation] Slitrkファミリータンパク質とIIa型受容体タンパク質チロシン脱リン酸化酵素がスプライスインサートに依存して相互作用するメカニズム (2015)
  • Author(s): 山形敦史、佐藤裕介、伊藤-後藤桜子、植村健、吉田知之、深井周也
  • Organizer: 第８８回日本生化学大会
  • Place of Presentation: 兵庫県神戸市 「神戸国際会議場」
  • Year and Date: 2015-12-03
• [Presentation] Insights into structure and assembly of GluD2-Cbln1-neurexin adhesion complex for cerebellar synapse formation (2015)
  • Author(s): Takeshi Uemura
  • Organizer: 第８８回日本生化学大会
  • Place of Presentation: 兵庫県神戸市 「神戸国際会議場」
  • Year and Date: 2015-12-01
  • Invited
• [Presentation] IIa 型受容体チロシンホスファターゼδ とインターロイキン１受容体タイプのシナプスオーガナイザー間の選択的スプラシング依存的相互作用制御の構造基盤 (2015)
  • Author(s): 山形敦史、吉田知之、佐藤裕介、伊藤-後藤桜子、植村健、森寿、三品昌美、深井周也
  • Organizer: 第８８回日本生化学大会
  • Place of Presentation: 兵庫県神戸市 「神戸国際会議場」
  • Year and Date: 2015-12-01
• [Presentation] 小脳シナプス形成を担うGluRδ2-Cbln1-neurexin接着分子複合体の構造基盤 (2015)
  • Author(s): 植村健
  • Organizer: 第53回日本生物物理学会年会
  • Place of Presentation: 石川県金沢市「金沢大学」
  • Year and Date: 2015-09-14
  • Invited
• [Presentation] Structural analysis of GluRδ2-Cbln1-Neurexin complex for cerebellar synapse formation (2015)
  • Author(s): Takeshi Uemura, Yusuke Sato, Atsushi Yamagata, Tomoyuki Yoshida, Sakurako Goto-Ito, Katsuhiko Tabuchi, Masayoshi Mishina, Shuya Fuka
  • Organizer: Neuro2015
  • Place of Presentation: 兵庫県神戸市 「神戸国際会議場」
  • Year and Date: 2015-07-30
• [Presentation] Structural basis of IL1RAPL1-PTPδ complex for neuronal synaptogenesis (2015)
  • Author(s): Tomoyuki Yoshida, Atsushi Yamagata, Yusuke Sato, Sakurako Ito:, Takeshi Uemura, Asami Maeda, Tomoko Shiroshima, Shiho Okamoto, Hisashi Mori, Masayoshi Mishina, Shuya Fukai
  • Organizer: Neuro2015
  • Place of Presentation: 兵庫県神戸市 「神戸国際会議場」
  • Year and Date: 2015-07-30
• [Presentation] tructural basis of the splicing dependent interaction between Slitrk2 and PTPδ (2015)
  • Author(s): Atsushi Yamagata, Yusuke Sato, Sakurako Goto-Ito, Takeshi Uemura, Asami Maeda, Tomoko Shiroshima, Tomoyuki Yoshida, Shuya Fuka
  • Organizer: Neuro2015
  • Place of Presentation: 兵庫県神戸市 「神戸国際会議場」
  • Year and Date: 2015-07-30
• [Presentation] C1qファミリー分子Cbln1とその受容体であるグルタミン酸受容体GluRδ2とneurexinによる小脳シナプス形成の調節 (2014)
  • Author(s): 植村 健
  • Organizer: 第37回日本分子生物学会年会
  • Place of Presentation: パシフィコ横浜（神奈川県横浜市）
  • Year and Date: 2014-11-26
  • Invited
• [Presentation] 線条体ドパミンD1受容体は文脈依存怖条件付け学習に必須である (2014)
  • Author(s): 三品昌美、池上賢、植村健、岸岡歩、崎村建司
  • Organizer: 第37回日本神経科学大会
  • Place of Presentation: パシフィコ横浜（神奈川県横浜市）
  • Year and Date: 2014-09-13
• [Presentation] The Atmospheric Scanning Electron Microscope (ASEM) Observes Axonal Segmentation and Synaptic Induction in Solution (2014)
  • Author(s): 2.佐藤 主税, 木下 貴明, 植村 健, 平野 和己, 川田 正晃, 西山英利, 佐藤 真理, 海老原 達彦, 須賀三雄, 西原祥子
  • Organizer: 2014 Microscopy & Microanalysis meeting
  • Place of Presentation: ハートフォード, アメリカ合衆国
  • Year and Date: 2014-08-05
• [Presentation] シナプス形成を司る GluRδ2-Cbln1-Neurexin三者複合体の構造解析 (2014)
  • Author(s): 植村 健
  • Organizer: 第14回日本蛋白質科学会年会
  • Place of Presentation: ワークピア横浜/横浜産貿ホール マリネリア（神奈川県横浜市）
  • Year and Date: 2014-06-27
  • Invited
• [Presentation] 受容体でコートした磁気ビーズによる初代培養シナプス形成誘導のASEM水中免疫電顕 (2014)
  • Author(s): 植村健、佐藤真理、西山英利、須賀三雄、佐藤主税
  • Organizer: 日本顕微鏡学会第70回記念学術講演会
  • Place of Presentation: 幕張メッセ国際会議場（千葉県千葉市）
  • Year and Date: 2014-05-11
• [Presentation] IL1RAPL1欠損マウスの行動学的解析 (2013)
  • Author(s): 安村美里, 吉田知之, 高雄啓三, 山崎真弥, 阿部学, 植村健, 宮川剛, 崎村建司, 三品昌美
  • Organizer: 第36回日本神経科学大会
  • Place of Presentation: 国立京都国際会館（京都府京都市）
  • Year and Date: 2013-06-21
• [Book] Synapse formation in the brain. Cortical Development. Neural Diversity and Neocortical Organization (2013)
  • Author(s): Mishina M, Yoshida T, Yasumura M, Uemura T
  • Total Pages: 282
  • Publisher: Springer Japan