Dual FRET技術を用いた長期神経可塑性機構の解読

• Project Area: Principles of memory dynamism elucidated from a diversity of learning systems
• Project/Area Number: 26115507
• 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: The University of Tokyo
• Principal Investigator: 尾藤 晴彦 東京大学, 医学(系)研究科(研究院), 教授 (00291964)
• Project Period (FY): 2014-04-01 – 2016-03-31
• Project Status: Completed (Fiscal Year 2015)
• Budget Amount: ¥9,360,000 (Direct Cost: ¥7,200,000、Indirect Cost: ¥2,160,000); Fiscal Year 2015: ¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000); Fiscal Year 2014: ¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000)
• Keywords: 神経科学 / シグナル伝達 / 脳神経

Outline of Annual Research Achievements

シナプスから核へのシグナリングの全貌を明らかにするため、前年度までの本研究では、樹状突起と細胞体の双方におけるカルシウム動態とリン酸化シグナル動態を同時測定する実験の準備を進めてきた。そこで、平成27年度においては、これらの成果をさらに発展させるため、in vivo マウス行動実験系にて確認する一連の実験を試みた。
具体的には、まず昨年度に開発した高速高感度カルシウムインディケーターR-CaMP2を用いた、in vivo 樹状突起イメージングの手法を開発・最適化し、プルキンエ細胞興奮に伴う樹状突起カルシウムシグナルの可視化に成功した（Gaffield et al. J. Neurophys. 2016）。さらに、記憶学習・報酬行動などに関与する投射線維の興奮を、同一投射線維内における多数の入力を細胞種別に記録する技術の開発を行った（Kim et al. Nature Met 2016）。これら技術開発と併行し、CaMKK-CaMKIVシグナルの細胞内局在を操作し、CREBリン酸化に対する影響を初代培養神経細胞系にて解明するとともに、このシグナル経路がin vivo記憶学習系にても、いかなる役割を果たしているのかを確認する実験を予備的に実施し、さらなる条件検討を行った。

Research Progress Status

27年度が最終年度であるため、記入しない。

Strategy for Future Research Activity

27年度が最終年度であるため、記入しない。

Research Products

• [Journal Article] Chronic imaging of Purkinje cell calcium activity in awake behaving mice. (2016)
  • Author(s): Gaffield MA, Amat SB, Bito H, Christie JM.
  • Journal Title: Journal of Neurophysiology Volume: 115 Issue: 1 Pages: 413-422
  • DOI: 10.1152/jn.00834.2015
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Simultaneous fast measurement of circuit dynamics at multiple sites across the mammalian brain. (2016)
  • Author(s): Kim CK, Yang SJ, Pichamoorthy N, Young NP, Kauvar I, Jennings JH, Lerner TN, Berndt A, Lee SY, Ramakrishnan C, Davidson TJ, Inoue M, Bito H, Deisseroth K.
  • Journal Title: Nature Methods Volume: 13 Issue: 4 Pages: 325-328
  • DOI: 10.1038/nmeth.3770
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] シナプスと記憶 ー シナプスと核の間の二方向性シグナリングの意義 (2016)
  • Author(s): 酒井一輝、金亮、奥野浩行、尾藤晴彦.
  • Journal Title: 生体の科学 Volume: 67 Pages: 4-8
• [Journal Article] Rational design of a high-affinity, fast, red calcium indicator R-CaMP2 (2015)
  • Author(s): Inoue M, Takeuchi A, Horigane S, Ohkura M, Gengyo-Ando K, Fujii H, Kamijo S, Takemoto-Kimura S, Kano M, Nakai J, Kitamura K, Bito H
  • Journal Title: Nat Methods Volume: 12 Issue: 1 Pages: 64
  • DOI: 10.1038/nmeth.3185
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Towards a better understanding of cognitive behaviors regulated by gene expression downstream of activity-dependent transcription factors (2014)
  • Author(s): Nonaka M, Kim R, Sharry S, Matsushima A, Takemoto-Kimura S, Bito H
  • Journal Title: Neurobiology of Learning and Memory Volume: 115 Pages: 21-29
  • DOI: 10.1016/j.nlm.2014.08.010
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Region-specific activation of CRTC1-CREB signaling mediates long-term fear memory (2014)
  • Author(s): Nonaka M, Kim R, Fukushima H, Sasaki K, Suzuki K, Okamura M, Ishii Y, Kawashima T, Kamijo S, Takemoto-Kimura S, Okuno H, Kida S, Bito H
  • Journal Title: Neuron Volume: 84 Issue: 1 Pages: 92-106
  • DOI: 10.1016/j.neuron.2014.08.049
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Presentation] An enduring cell labeling for active cell ensembles within a restricted time window. (2015)
  • Author(s): Araki A, Imayoshi I, Minatohara K, Kim R, Kawashima T, Kageyama R, Bito H, Okuno H.
  • Organizer: 第38回日本分子生物学会・第88回日本生化学会大会合同大会
  • Place of Presentation: 神戸ポートアイランド(神戸市)
  • Year and Date: 2015-12-01
• [Presentation] Rational design of ultrafast, high-affinity red calcium indicator for monitoring neuronal activity. (2015)
  • Author(s): Inoue M, Takeuchi A, Horigane S, Fujii H, Kamijo S, Takemoto-Kimura S, Ohkura M, Gengyo-Ando K, Kano M, Nakai J, Kitamura K, Bito H.
  • Organizer: The 45th Annual Meeting of the Society for Neuroscience
  • Place of Presentation: Chicago, USA
  • Year and Date: 2015-10-17
  • Int'l Joint Research
• [Presentation] Arc/Arg3.1 knockout mice exhibit impaired behavioral flexibility in a serial reversal learning task (2015)
  • Author(s): Okuno H, Endo T, Kim R, MInatohara K, Araki A, Abe M,Kakeyama M, Sakimura K, Bito H.
  • Organizer: The 45th Annual Meeting of the Society for Neuroscience
  • Place of Presentation: Chicago, USA
  • Year and Date: 2015-10-17
  • Int'l Joint Research
• [Presentation] Synapse-to-Nucleus Signaling and CREB Regulation in Fear Memory. (2015)
  • Author(s): Bito H.
  • Organizer: Gordon Research Conference on Amygdala in Health & Disease.
  • Place of Presentation: Stonhill College, Easton, Massachussetts, USA.
  • Year and Date: 2015-08-02
  • Int'l Joint Research / Invited
• [Presentation] 高感度、超高速赤色カルシウムインディケーターの合理的設計による開発. (2015)
  • Author(s): 井上昌俊, 竹内敦也, 堀金慎一郎, 大倉正道, 安藤恵子, 藤井哉, 上條諭志, 竹本-木村 さやか, 狩野方伸, 中井淳一, 喜多村和郎, 尾藤晴彦.
  • Organizer: 第38回日本神経科学大会
  • Place of Presentation: 神戸国際会議場・神戸国際展示場（神戸）
  • Year and Date: 2015-07-28
• [Presentation] Understanding Arc to understand a memory engram. (2015)
  • Author(s): Bito H.
  • Organizer: The 28th Annual Meeting on the Neurobiology of Learning and Memory.
  • Place of Presentation: University of California at Irvine, Irvine, California, USA.
  • Year and Date: 2015-04-23
  • Int'l Joint Research / Invited
• [Presentation] Activity-dependent gene expression in health and diseases. (2014)
  • Author(s): Bito H
  • Organizer: 第36回日本生物学的精神医学会 第57回日本神経化学会合同大会
  • Place of Presentation: 奈良
  • Year and Date: 2014-09-28 – 2014-09-30
  • Invited
• [Presentation] Activity-dependent Arc expression: mechanism, function and application. (2014)
  • Author(s): Bito H
  • Organizer: 4th Leibniz Institute of Neurobiology Symposium
  • Place of Presentation: Tnagermuende, Germany
  • Year and Date: 2014-08-25 – 2014-08-28
  • Invited