精神神経疾患を理解するシナプスのスクラップ＆ビルド

• Project Area: Dynamic regulation of brain function by Scrap & Build system
• Project/Area Number: 16H06463
• Research Category: Grant-in-Aid for Scientific Research on Innovative Areas (Research in a proposed research area)
• Allocation Type: Single-year Grants
• Review Section: Biological Sciences
• Research Institution: Institute of Physical and Chemical Research
• Principal Investigator: 内匠 透 国立研究開発法人理化学研究所, 脳神経科学研究センター, チームリーダー (00222092)
• Co-Investigator(Kenkyū-buntansha): 萬代 研二 北里大学, 医学部, 教授 (50322186)
• Project Period (FY): 2016-06-30 – 2021-03-31
• Project Status: Granted (Fiscal Year 2019)
• Budget Amount: ¥99,190,000 (Direct Cost: ¥76,300,000、Indirect Cost: ¥22,890,000); Fiscal Year 2019: ¥18,980,000 (Direct Cost: ¥14,600,000、Indirect Cost: ¥4,380,000); Fiscal Year 2018: ¥18,460,000 (Direct Cost: ¥14,200,000、Indirect Cost: ¥4,260,000); Fiscal Year 2017: ¥18,980,000 (Direct Cost: ¥14,600,000、Indirect Cost: ¥4,380,000); Fiscal Year 2016: ¥23,790,000 (Direct Cost: ¥18,300,000、Indirect Cost: ¥5,490,000)
• Keywords: シナプス / スパイン / モデルマウス

Outline of Annual Research Achievements

興奮性シナプスは、神経細胞樹状突起から突き出たサブミクロンの突起（スパイン）上に形成される。スパインの数は形成（ビルド）と破壊（スクラップ）の動的平衡によって決まり、発達期に増加し成長後にゆるやかに減少することが知られているがその分子基盤は未解明である。我々は自閉スペクトラム症（ASD）モデルマウスに共通する所見として大脳皮質の錐体細胞でのスパイン形成と破壊が共に亢進することを見出した。本研究では、自閉症原因遺伝子とその下流に焦点を当て、大脳皮質でのスクラップ＆ビルド機構の分子機序を明らかにし、行動解析やin vivoスパインイメージング法によってスクラップ＆ビルド現象の破綻と高次機能障害との関係を解明する。
自閉症モデルマウス（patDp/+）のスパイン動態の変容(大脳皮質の錐体細胞でのスパイン形成と破壊が共に亢進すること)に関わる分子として、２光子顕微鏡を用いたスパインのin vivoイメージング法を用いて、ヒト１５番染色体（15q11-13）父性重複領域に存在するNdnを同定した。またpatDp/+マウスのシナプトソームを用いたプロテオミクス解析を行うことにより、スパイン動態の変容に関わる蛋白質群の候補を得た。これらの結果から、スパイン動態と神経細胞の局所翻訳の連関が示唆された。
一方、シナプス・細胞接着に関する分子の機能解析では、自閉症患者のエクソームシーケンス解析から新規Neuroligin1自閉症変異を同定した。さらにCRISPR/Cas9法により患者変異を有する点変異ノックインマウスを作製した。

Current Status of Research Progress

2: Research has progressed on the whole more than it was originally planned.

Reason

自閉症モデルマウスのスパイン動態の変容に関わる分子として、自閉症様行動がmatDp/+マウスではなくpatDp/+マウスに見られることから、父性発現遺伝子群（Mkrn3, Magel2, Ndn, Snrpn）を子宮内穿孔法により遺伝子導入することにより、大脳皮質に発現させた。これらのマウスの２光子顕微鏡によるスパインのin vivoイメージングをしたところ、父性発現遺伝子群のうちNdnを発現させた際にpatDp/+マウスのスパイン表現型と似た表現型を得たことにより、Ndnがその原因遺伝子として注目された。一方、patDp/+マウスのシナプトソームを用いたプロテオミクス解析の結果、タンパク合成に関わる分子が同定され、局所翻訳とスパイン動態のリンクが示唆された。

Strategy for Future Research Activity

自閉症患者のエクソームシーケンス解析により、シナプス形成に関わる分子として、新規Neuroligin1自閉症変異を見いだした。CRISPR/Cas9系を用いて本自閉症変異ノックインマウスを既に作製している。今後は本変異体のin vitro解析、さらに本モデルマウスの行動解析を含めた多面的解析を行う。

Research Products

• [Journal Article] CHRONO integrates behavioral stress and epigenetic control of metabolism. (2017)
  • Author(s): Hatanaka F, Takumi T
  • Journal Title: Ann Med Volume: 49 Pages: 352-356
  • DOI: 10.1080/07853890.2016.1276301
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Network Dynamics Mediate Circadian Clock Plasticity (2017)
  • Author(s): Azzi A, Evans JA, Leise T, Myung J, Takumi T, Davidson AJ, Brown SA
  • Journal Title: Neuron Volume: 93 Pages: 441-450
  • DOI: 10.1016/j.neuron.2016.12.022
  • Peer Reviewed / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Ror2 signaling regulates Golgi structure and transport through IFT20 for tumor invasiveness. (2017)
  • Author(s): Nishita, M., Park, S.-Y., Nishio, T., Kamizaki, K., Wang, Z., Tamada, K., Takumi, T., Hashimoto, R., Otani, H., Pazour, G.J., Hsu, V.W., and Minami, Y.
  • Journal Title: Sci Rep. Volume: 7 Pages: 1-1
  • DOI: 10.1038/s41598-016-0028-x
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Distinct Defects in Spine Formation or Pruning in Two Gene Duplication Mouse Models of Autism. (2017)
  • Author(s): Wang M, Li H, Takumi T, Qiu Z, Xu X, Yu X, Bian WJ
  • Journal Title: Neurosci Bull Volume: 33 Pages: 143-152
  • DOI: 10.1007/s12264-017-0111-8
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Rodent models of genetic and chromosomal variations in psychiatric disorders. (2017)
  • Author(s): Nomura J, Kannan G, Takumi T
  • Journal Title: Psychiatry Clin Neurosci Volume: - Pages: 508-517
  • DOI: 10.1111/pcn.12524
  • Peer Reviewed / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Behavioral and neuroanatomical analyses in a genetic mouse model of 2q13 duplication (2017)
  • Author(s): Kishimoto K, Nomura J, Ellegood J, Fukumoto K, Lerch JP, Moreno-De-Luca D, Bourgeron T, Tamada K, Takumi T
  • Journal Title: Genes to Cells Volume: 22 Pages: 436-451
  • DOI: 10.1111/gtc.12487
  • Peer Reviewed / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] p53 induces formation of NEAT1 lncRNA-containing paraspeckles that modulate replication stress response and chemosensitivity. (2017)
  • Author(s): Chujo T, Yamazaki T, Kawaguchi T, Kurosaka S, Takumi T, Nakagawa S, Hirose T
  • Journal Title: Nat Med Volume: 36 Pages: 1447-1462
  • DOI: 10.15252/embj.201695848
  • Peer Reviewed / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Serotonin rebalances cortical tuning and behavior linked autism symptoms in 15q11-13 CNV mice. (2017)
  • Author(s): Nakai N, Nagano M, Saitow F, Watanabe Y, Kawamura Y, Kawamoto A, Tamada K, Mizuma H, Onoe H, Watanabe Y, Monai H, Hirase H, Nakatani J, Inagaki H, Kawada T, Miyazaki T, Watanabe M, Sato Y, Okabe S, Kitamura K, Kano M, Hashimoto K, Suzuki H, Takumi T
  • Journal Title: Sci Adv Volume: -
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Roles of afadin in the formation of the cellular architecture of the mouse hippocampus and dentate gyrus. (2017)
  • Author(s): Miyata M, Maruo T, Kaito A, Wang S, Yamamoto H, Fujiwara T, Mizoguchi A, Mandai K, Takai Y
  • Journal Title: Molecular and Cellular Neuroscience Volume: 79 Pages: 34-44
  • DOI: 10.1016/j.mcn.2016.12.007
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Aging-dependent expression of synapse-related proteins in the mouse brain (2017)
  • Author(s): Shiotani H, Maruo T, Sakakibara S, Miyata M, Mandai K, Mochizuki H, Takai Y
  • Journal Title: Genes to Cells Volume: 22 Pages: 472-484
  • DOI: 10.1111/gtc.12489
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Multiple roles of afadin in the ultrastructural morphogenesis of mouse hippocampal mossy fiber synapses (2017)
  • Author(s): Sai K, Wang S, Kaito A, Fujiwara T, Maruo T, Itoh Y, Miyata M, Sakakibara S, Miyazaki N, Murata K, Yamaguchi Y, Haruta T, Nishioka H, Motojima Y, Komura M, Kimura K, Mandai K, Takai Y, Mizoguchi A.
  • Journal Title: Journal of Comparative Neurology Volume: - Pages: 2719-2734
  • DOI: 10.1002/cne.24238
  • Peer Reviewed
• [Journal Article] Structural, super-resolution microscopy analysis of paraspeckle nuclear body organization (2016)
  • Author(s): Wu M, Takemoto M, Taniguchi M, Takumi T, Okazaki T, Song WJ
  • Journal Title: Journal of Cell Biology Volume: 311 Pages: 817-830
  • DOI: 10.1083/jcb.201601071
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] CHD8 haploinsufficiency results in autistic-like phenotypes in mice. (2016)
  • Author(s): Katayama Y, Nishiyama M, Shoji H, Ohkawa Y, Kawamura A, Sato T, Suyama M, Takumi T, Miyakawa T, Nakayama KI.
  • Journal Title: Nature Volume: 537 Issue: 7622 Pages: 28636-28636
  • DOI: 10.1038/nature19357
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Exome sequencing in the knockin mice generated using the CRISPR/Cas system (2016)
  • Author(s): Nakajima K, Kazuno AA, Kelsoe J, Nakanishi M, Takumi T, Kato T
  • Journal Title: Scientific Reports Volume: 6 Pages: 34703-34703
  • DOI: 10.1038/srep34703
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] FUS/TLS acts as an aggregation-dependent modifier of polyglutamine disease model mice. (2016)
  • Author(s): Kino Y, Washizu C, Kurosawa M, Yamada M, Doi H, Takumi T, Adachi H, Katsuno M, Sobue G, Hicks GG, Hattori N, Shimogori T, Nukina N.
  • Journal Title: Scientific Reports Volume: 6 Pages: 35236-35236
  • DOI: 10.1038/srep35236
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Localization of nectin-2delta at perivascular astrocytic endfoot processes and degeneration of astrocytes and neurons in nectin-2 knockout mouse brain. (2016)
  • Author(s): Miyata M, Mandai K, Maruo T, Sato J, Shiotani H, Kaito A, Itoh Y, Wang S, Fujiwara T, Mizoguchi A, Takai Y, Rikitake Y
  • Journal Title: Brain Research Volume: 1649 Pages: 90-101
  • DOI: 10.1016/j.brainres.2016.08.023
  • Peer Reviewed / Acknowledgement Compliant
• [Presentation] Necdin regulates spine dynamics in 15q duplication mode mice (2017)
  • Author(s): 戸谷豪、福本景太、玉田紘太、田中慎二、三澤日出巳、岡部繁男、内匠透
  • Organizer: 第１２２回日本解剖学会全国学術集会
  • Place of Presentation: 長崎大学、長崎
  • Year and Date: 2017-03-28
  • Invited
• [Presentation] Abnormal synapse dynamics in mouse models for autism spectrum disorders (2017)
  • Author(s): Shinji Tanaka, Masaaki Isshiki, Risa Iguchi, Shinji Urata, Shunsuke Mizutani, Toshihiko Kuriu, Katsuhiko Tabuchi, Toru Takumi, Shigeo Okabe
  • Organizer: 第１２２回日本解剖学会全国学術集会
  • Place of Presentation: 長崎大学、長崎
  • Year and Date: 2017-03-28
  • Invited
• [Presentation] Copy number variation models with autism (2016)
  • Author(s): Toru Takumi
  • Organizer: 14th Meeting of the Asian-Pacific Society for Neurochemistry
  • Place of Presentation: Kuala Lumpur, Malaysia
  • Year and Date: 2016-08-29
  • Int'l Joint Research / Invited
• [Presentation] Modeling autism (2016)
  • Author(s): Toru Takumi
  • Organizer: 第３９回日本神経科学大会
  • Place of Presentation: パシフィコ横浜、横浜
  • Year and Date: 2016-07-22
  • Invited
• [Presentation] Homozygotes of model mice for human chromosome 15q11-q13 duplication show severe developmental abnormalities. (2016)
  • Author(s): Jin Nakatani, Natsu Koyama, Seiji Hitoshi, Toru Takumi, Tomoko Kato, Ikuo Tooyama, Akihiko Shiino, Shigehiro Morikawa, Toshiro Inubushi
  • Organizer: 第３９回日本神経科学大会
  • Place of Presentation: パシフィコ横浜、横浜
  • Year and Date: 2016-07-20
• [Presentation] Neurobehavioral abnormalities in a mouse model for huma 15q25.2-25.3 deletion (2016)
  • Author(s): Jun Nomura, Akifumi Kanda, Yusuke Sotomaru, Toru Takumi
  • Organizer: 第３９回日本神経科学大会
  • Place of Presentation: パシフィコ横浜、横浜
  • Year and Date: 2016-07-20
• [Presentation] Dosage-dependent cognitive dysfunctions in a genetic mouse model of 2q13 (Nphp1) duplication (2016)
  • Author(s): Keiko Kishimoto, Jun Nomura, Kota Tamada, Thomas Bourgeron, Mereno Du Luca Daniel, Toru Takumi
  • Organizer: 第３９回日本神経科学大会
  • Place of Presentation: パシフィコ横浜、横浜
  • Year and Date: 2016-07-20
• [Presentation] Identification and functional characterization of a novel Neuroligin1 variant in autism spectrum disorder (2016)
  • Author(s): Moe Nakanishi, Jun Nomura, Xiaoxi Liu, Takashi Arai, Eiki Takahashi, Maja Bucan, Li Zhou, Manabu Abe, Kenji Sakimura, Toru Takumi
  • Organizer: 第３９回日本神経科学大会
  • Place of Presentation: パシフィコ横浜、横浜
  • Year and Date: 2016-07-20
• [Presentation] Phenotypic analysis of a mouse model for 15q25.2-25.3 deletion syndrome (2016)
  • Author(s): Jun Nomura, Akifumi Kanda, Yusuke Sotomaru, Toru Takumui
  • Organizer: 30th CINP World Congress of Neuropsychopharmacology
  • Place of Presentation: Seoul, South Korea
  • Year and Date: 2016-07-03
  • Int'l Joint Research
• [Presentation] A novel mutation associated autism in Neuroligin1 (2016)
  • Author(s): Moe Nakanishi, Takashi Arai, Maja Bucan, Xiao Ji, Xiaoxi Liu, Jun Nomura, Eiki Takahashi, Toru Takumi
  • Organizer: 30th CINP World Congress of Neuropsychopharmacology
  • Place of Presentation: Seoul, South Korea
  • Year and Date: 2016-07-03
  • Int'l Joint Research
• [Presentation] 自閉スペクトラム症の生物学的理解 (2016)
  • Author(s): 内匠透
  • Organizer: 第４６回日本神経精神薬理学会年会
  • Place of Presentation: Seoul, South Korea
  • Year and Date: 2016-07-02
  • Invited
• [Presentation] 発達障害と相関する新規CNV(15q25.2-25.3)動物モデルの開発 (2016)
  • Author(s): 野村淳、神田暁史、外丸祐介、内匠透
  • Organizer: 第４６回日本神経精神薬理学会年会
  • Place of Presentation: Seoul, South Korea
  • Year and Date: 2016-07-02
• [Presentation] Neuroligin1における自閉症関連変異の同定と機能解析 (2016)
  • Author(s): 仲西萌絵、野村淳、Xiaoxi Liu、Maja Bucan、高橋英機、阿部学、li Zhou、崎村健司、内匠透
  • Organizer: 第４６回日本神経精神薬理学会年会
  • Place of Presentation: Seoul, South Korea
  • Year and Date: 2016-07-02