Molecular mechanism of synapse maturation and pathophysiology of epilepsy

• Project/Area Number: 20H00459
• Research Category: Grant-in-Aid for Scientific Research (A)
• Allocation Type: Single-year Grants
• Section: 一般
• Review Section: Medium-sized Section 43:Biology at molecular to cellular levels, and related fields
• Research Institution: National Institute for Physiological Sciences
• Principal Investigator: Fukata Masaki 生理学研究所, 分子細胞生理研究領域, 教授 (00335027)
• Project Period (FY): 2020-04-01 – 2023-03-31
• Project Status: Completed (Fiscal Year 2022)
• Budget Amount: ¥45,240,000 (Direct Cost: ¥34,800,000、Indirect Cost: ¥10,440,000); Fiscal Year 2022: ¥14,040,000 (Direct Cost: ¥10,800,000、Indirect Cost: ¥3,240,000); Fiscal Year 2021: ¥14,820,000 (Direct Cost: ¥11,400,000、Indirect Cost: ¥3,420,000); Fiscal Year 2020: ¥16,380,000 (Direct Cost: ¥12,600,000、Indirect Cost: ¥3,780,000)
• Keywords: シナプス / タンパク質 / 脂質 / 脳・神経 / ナノドメイン / ナノカラム / てんかん / 酵素 / パルミトイル化

Outline of Research at the Start

脳の生理と病態を理解するには、神経細胞間の情報伝達を担う“シナプス”の理解が必要不可欠である。シナプス伝達の効率は常に一定ではなく、外界刺激によって柔軟に変化し、これが記憶や学習などの脳高次機能の基盤となる。本研究では、申請者らが独自に見出した (1)パルミトイル化脂質修飾関連酵素と(2)分泌リガンド・受容体LGI1-ADAM22を起点として、シナプスの成熟機構とその破綻によるシナプス疾患の分子病態を解明する。

Outline of Final Research Achievements

To understand the physiology and pathology of the brain, it is essential to understand synapses, which are responsible for information transfer between neurons. Recently, many proteins involved in synapse formation have been reported, but the process by which immature synapses mature into functional synapses is not well understood. Here, we found that the LGI1-ADAM22-PSD-95 protein complex elaborately regulates synaptic transmission by aligning the nanodomains in the pre- and post-synapse. In addition, we found that disruption of the LGI1-ADAM22-PSD-95 pathway causes epileptic encephalopathy in humans and mice.

Academic Significance and Societal Importance of the Research Achievements

従来の光学顕微鏡や電子顕微鏡を用いた解析では、シナプス内部やシナプス間隙をまたぐ分子の相互配置や動的変化に迫ることは困難であった。本研究では、超解像イメージング法や脳疾患モデルを用いて、シナプス内部のタンパク質の配置メカニズムを明らかにした。また、シナプス疾患をシナプスの形態や数の異常という従来の観点を超えて、シナプス-ナノドメインの構造・機能的変容として捉えた点も意義深い。

Research Products

• [Int'l Joint Research] UCL/The University of Edinburgh(英国)
• [Int'l Joint Research] University of Helsinki(フィンランド)
• [Int'l Joint Research] Erasmus University/Leiden University(オランダ)
• [Int'l Joint Research] DZNE/Charite-Universitatsmedizin(ドイツ)
• [Int'l Joint Research] Charles University in Prague(チェコ)
• [Int'l Joint Research]
• [Int'l Joint Research] UCSF(米国)
• [Int'l Joint Research] UCL/The University of Edinburgh(英国)
• [Int'l Joint Research] DZNE/Charite-Universitatsmedizin(ドイツ)
• [Int'l Joint Research] Aarhus University(デンマーク)
• [Int'l Joint Research] Erasmus University/Leiden University(オランダ)
• [Int'l Joint Research]
• [Int'l Joint Research] UCSF/UCSD(米国)
• [Int'l Joint Research] DZNE(ドイツ)
• [Int'l Joint Research] IST(オーストリア)
• [Int'l Joint Research] UCL/The University of Edinburgh(英国)
• [Int'l Joint Research] University of Helsinki(フィンランド)
• [Journal Article] Anti-LGI4 antibody is a novel juxtaparanodal autoantibody for chronic inflammatory demyelinating polyneuropathy (2023)
  • Author(s): Zhang X, Kira J, Ogata H, Imamura T, Mitsuishi M, Fujii T, Kobayashi M, Kitagawa K, Namihira Y, Ohya Y, Maimaitijiang G, Yamasaki R, Fukata Y, Fukata M, Isobe N, Nakamura Y.
  • Journal Title: Neurology Neuroimmunology and Neuroinflammation Volume: 10(2) Issue: 2 Pages: 1-13
  • DOI: 10.1212/nxi.0000000000200081
  • Peer Reviewed / Open Access
• [Journal Article] S-Palmitoylation of Tyrosinase at Cysteine500 Regulates Melanogenesis (2023)
  • Author(s): Niki Yoko、Adachi Naoko、Fukata Masaki、Fukata Yuko、Oku Shinichiro、Makino-Okamura Chieko、Takeuchi Seiji、Wakamatsu Kazumasa、Ito Shosuke、Declercq Lieve、Yarosh Daniel B.、Mammone Tomas、Nishigori Chikako、Saito Naoaki、Ueyama Takehiko
  • Journal Title: Journal of Investigative Dermatology Volume: 143 Issue: 2 Pages: 317-327.e6
  • DOI: 10.1016/j.jid.2022.08.040
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] てんかん発症を抑制するためのLGI1-ADAM22タンパク質複合体の量的制御機構 (2023)
  • Author(s): 横井紀彦、深田優子、深田正紀
  • Journal Title: 生化学 Volume: -
  • Peer Reviewed
• [Journal Article] Biallelic ADAM22 pathogenic variants cause progressive encephalopathy and infantile-onset refractory epilepsy (2022)
  • Author(s): van der Knoop M Marieke M*、Maroofian Reza*、Fukata Yuko*、van Ierland Yvette、Karimiani G Ehsan、Lehesjoki Anna-Elina、Muona Mikko、Paetau Anders、Miyazaki Yuri、Hirano Yoko、Selim Laila、-----32名-----、Striano Pasquale、Severino Mariasavina、Fukata Masaki#、Hilhorst-Hofstee Yvonne#、Houlden Henry#
  • Journal Title: Brain Volume: - Issue: 7 Pages: 2301-2312
  • DOI: 10.1093/brain/awac116
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Palmitoylation of the small GTPase Cdc42 by DHHC5 modulates spine formation and gene transcription (2022)
  • Author(s): Wirth Alexander、Labus Josephine、Abdel Galil Dalia、Schill Yvonne、Schmidt Silke、Bunke Tania、Gorinski Nataliya、Yokoi Norihiko、Fukata Masaki、Ponimaskin Evgeni
  • Journal Title: Journal of Biological Chemistry Volume: 298 Issue: 6 Pages: 102048-102048
  • DOI: 10.1016/j.jbc.2022.102048
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Insight into the function of a unique voltage-sensor protein (TMEM266) and its short form in mouse cerebellum (2022)
  • Author(s): Kawai T、Narita H、Konno K、Akter S、Andriani RT、Iwasaki H、Nishikawa S、Yokoi N、Fukata Y、Fukata M、Wiriyasermkul P、Kongpracha P、Nagamori S、Takao K、Miyakawa T、Abe M、Sakimura K、Watanabe M、Nakagawa A、Okamura Y
  • Journal Title: Biochemical Journal Volume: 479 Issue: 11 Pages: 1127-1145
  • DOI: 10.1042/bcj20220033
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] MAGUKs are essential, but redundant, in long-term potentiation (2021)
  • Author(s): Chen Xiumin、Fukata Yuko、Fukata Masaki、Nicoll Roger A.
  • Journal Title: Proceedings of the National Academy of Sciences Volume: 118 Issue: 28
  • DOI: 10.1073/pnas.2107585118
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Trans-synaptic LGI1-ADAM22?MAGUK in AMPA and NMDA receptor regulation (2021)
  • Author(s): Fukata Yuko、Hirano Yoko、Miyazaki Yuri、Yokoi Norihiko、Fukata Masaki
  • Journal Title: Neuropharmacology Volume: 194 Pages: 108628-108628
  • DOI: 10.1016/j.neuropharm.2021.108628
  • Peer Reviewed / Open Access
• [Journal Article] 14-3-3 proteins stabilize LGI1-ADAM22 levels to regulate seizure thresholds in mice (2021)
  • Author(s): Norihiko Yokoi, Yuko Fukata, Kei Okatsu, Atsushi Yamagata, Yan Liu , Makoto Sanbo, Yuri Miyazaki, Teppei Goto, Manabu Abe , Hidetoshi Kassai, Kenji Sakimura, Dies Meijer, Masumi Hirabayashi, Shuya Fukai, Masaki Fukata
  • Journal Title: Cell Reports Volume: 37 Issue: 11 Pages: 110107-110107
  • DOI: 10.1016/j.celrep.2021.110107
  • NAID: 120007177780
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] A novel red fluorescence dopamine biosensor selectively detects dopamine in the presence of norepinephrine in vitro (2021)
  • Author(s): Nakamoto Chihiro、Goto Yuhei、Tomizawa Yoko、Fukata Yuko、Fukata Masaki、Harpsoe Kasper、Gloriam David E.、Aoki Kazuhiro、Takeuchi Tomonori
  • Journal Title: Molecular Brain Volume: 14 Issue: 1 Pages: 173-173
  • DOI: 10.1186/s13041-021-00882-8
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Encephalitis patient-derived monoclonal GABAa receptor antibodies cause epileptic seizures (2021)
  • Author(s): Kreye Jakob、Wright K Sukhvir、van Casteren Adriana、Stoffler Laura、Machule Marie-Luise、Reincke S Momsen、Nikolaus Marc、van Hoof Scott、Sanchez-Sendin Elisa、Homeyer A Marie、Cordero Gomez Cesar、Kornau Hans-Christian、-----12名-----、Wickel Jonathan、Geis Christian、Fukata Yuko、Fukata Masaki、Pruss Harald
  • Journal Title: Journal of Experimental Medicine Volume: 218
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] LGI1-ADAM22-MAGUK configures transsynaptic nanoalignment for synaptic transmission and epilepsy prevention (2021)
  • Author(s): Fukata Yuko、Chen Xiumin、Chiken Satomi、Hirano Yoko、Yamagata Atsushi、Inahashi Hiroki、Sanbo Makoto、Sano Hiromi、Goto Teppei、Hirabayashi Masumi、Kornau Hans-Christian、Pruss Harald、Nambu Atsushi、Fukai Shuya、Nicoll Roger A.、Fukata Masaki
  • Journal Title: Proceedings of the National Academy of Sciences Volume: 118 Issue: 3 Pages: 1-12
  • DOI: 10.1073/pnas.2022580118
  • NAID: 120006976050
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Canonical versus non-canonical transsynaptic signaling of neuroligin 3 tunes development of sociality in mice (2021)
  • Author(s): Yoshida T, Yamagata A, Imai A, Kim J, Izumi H, Nakashima S, Shiroshima T, Maeda A, Iwasawa-Okamoto S, Azechi K, Osaka F, Saitoh T, Maenaka K, Shimada T, Fukata Y, Fukata M, Matsumoto J, Nishijo H, Takao K, Tanaka S, Okabe S, Tabuchi K, Uemura T, Mishina M, Mori H & Fukai S
  • Journal Title: Nature Communications Volume: 12(1) Issue: 1 Pages: 1848-1848
  • DOI: 10.1038/s41467-021-22059-6
  • NAID: 120006996978
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Encephalitis patient derived monoclonal GABAA receptor antibodies cause catatonia and epileptic seizures (2021)
  • Author(s): Kreye Jakob、Wright Sukhvir K.、van Casteren Adriana、Machule Marie-Luise、Reincke S. Momsen、Nikolaus Marc、Stoffler Laura、van Hoof Scott、Sanchez-Sendin Elisa、Kornau Hans-Christian、Kaindl Angela M.、Wilson Max A.、Greenhill Stuart、他5名、Geis Christian、Fukata Yuko、Fukata Masaki、Pruss Harald
  • Journal Title: bioRxiv Volume: -
  • DOI: 10.1101/2021.01.28.428602
  • Open Access / Int'l Joint Research
• [Journal Article] APEGS法によるタンパク質パルミトイル化修飾の定量 (2021)
  • Author(s): 深田正紀、横井紀彦、深田優子
  • Journal Title: 電気泳動 Volume: 65
  • NAID: 130008072916
  • Peer Reviewed
• [Journal Article] Forelimb movements evoked by optogenetic stimulation of the macaque motor cortex (2020)
  • Author(s): Watanabe H, Sano H, Chiken S, Kobayashi K, Fukata Y, Fukata M, Mushiake H, Nambu A
  • Journal Title: Nat Commun Volume: 11 Issue: 1 Pages: 3253-3253
  • DOI: 10.1038/s41467-020-16883-5
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] A genetically encoded red fluorescence dopamine biosensor enables dual imaging of dopamine and norepinephrine (2020)
  • Author(s): Nakamoto Chihiro、Goto Yuhei、Tomizawa Yoko、Fukata Yuko、Fukata Masaki、Harpsoe Kasper、Gloriam David E.、Aoki Kazuhiro、Takeuchi Tomonori
  • Journal Title: bioRxiv Volume: -
  • DOI: 10.1101/2020.05.25.115162
  • Open Access / Int'l Joint Research
• [Presentation] シナプスのナノ構築機構と病態 (2023)
  • Author(s): 深田優子、横井紀彦、宮﨑裕理、深田正紀
  • Organizer: 第128回日本解剖学会総会・全国学術集会
  • Invited
• [Presentation] Function and regulation of ADAM22, a receptor for LGI1, in epileptic disorders (2022)
  • Author(s): Fukata M, Fukata Y
  • Organizer: SYNABS & CONNECT-GENERATE Symposium 2022
  • Int'l Joint Research / Invited
• [Presentation] 14-3-3 proteins stabilize LGI1-ADAM22 levels to prevent epilepsy in mice (2022)
  • Author(s): Yokoi N, Fukata Y, Okatsu K, Yamagata A, Liu Y, Sanbo M, Miyazaki Y, Goto T, Hirabayashi M, Fukai S, Fukata M
  • Organizer: The 45th annual meeting of the Japan Neuroscience Society
• [Presentation] シナプス機能と脳の疾患に関する研究. 第 4 回 FUJITA ブレインサイエンスセミナー (2022)
  • Author(s): 深田正紀
  • Organizer: 第 4 回 FUJITA ブレインサイエンスセミナー
  • Invited
• [Presentation] Proteomic characterization of ABHD17B, a depalmitoylating enzyme, in the mouse brain (2022)
  • Author(s): Egawa T, Hirata T, Yokoi N, Miyazaki Y, Sanbo M, Goto T, Inahashi H, Kobayashi K, Hirabayashi M, Fukata Y, Fukata M
  • Organizer: 第95回日本生化学大会
• [Presentation] Trans-synaptic LGI1-ADAM22-MAGUK protein complex in synaptic functions and disorders (2021)
  • Author(s): Fukata Masaki、Fukata Yuko
  • Organizer: SYNABS Symposium (Germany, Virtual meeting)
  • Int'l Joint Research / Invited
• [Presentation] Acyl-PEGyl exchange gel-shift (APEGS) assay for palmitoylation quantification (2020)
  • Author(s): Fukata Masaki、Yokoi Norihiko、Fukata Yuko
  • Organizer: 第71回 日本電気泳動学会総会
  • Invited
• [Presentation] Synaptic function regulated by palmitate cycling on PSD-95 and trans-synaptic LGI1 and ADAM22 (2020)
  • Author(s): Fukata Masaki、Yuri Miyazaki、Yokoi Norihiko、Fukata Yuko
  • Organizer: 第43回日本分子生物学会年会
  • Invited
• [Book] 膜タンパク質工学ハンドブック (2020)
  • Author(s): 平野瑶子、宮﨑裕理、横井紀彦、平田哲也、深田正紀、深田優子
  • Total Pages: 17
  • Publisher: エヌ・ティー・エス
  • ISBN: 9784860435370
• [Remarks] 研究室ホームページ
  • URL: https://www.nips.ac.jp/fukata/
• [Remarks] 研究報告_生理研ホームページ
  • URL: https://www.nips.ac.jp/nips_research/2022/04/_adam22_17.html
• [Remarks] プレスリリース 新たなてんかん治療戦略を提案
  • URL: https://www.nips.ac.jp/release/2021/12/_adam22.html
• [Remarks] プレスリリース 新たなてんかん治療戦略を提案
  • URL: https://www.amed.go.jp/news/release_20211215.html
• [Remarks] プレスリリース 新規赤色蛍光ドーパミンバイオセンサーの開発
  • URL: https://www.nibb.ac.jp/press/2021/12/20.html
• [Remarks] プレスリリース：神経シナプス間の軸調整を担う分子群を発見―新たなてんかん病態を解明―
  • URL: https://www.nips.ac.jp/release/2021/01/post_426.html