Development and application of a toolkit to visualize/manipulate neural activity using E-SARE and next-generation GECIs

• Project/Area Number: 17K19442
• Research Category: Grant-in-Aid for Challenging Research (Exploratory)
• Allocation Type: Multi-year Fund
• Research Field: Neuroscience and related fields
• Research Institution: The University of Tokyo
• Principal Investigator: BITO Haruhiko 東京大学, 大学院医学系研究科(医学部), 教授 (00291964)
• Project Period (FY): 2017-06-30 – 2019-03-31
• Project Status: Completed (Fiscal Year 2018)
• Budget Amount: ¥6,370,000 (Direct Cost: ¥4,900,000、Indirect Cost: ¥1,470,000); Fiscal Year 2018: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000); Fiscal Year 2017: ¥4,420,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥1,020,000)
• Keywords: 神経科学 / 脳・神経 / シグナル伝達 / 神経活動

Outline of Final Research Achievements

We constructed an experimental system that simultaneously expresses multi-color next-generation GECI probes in a cell type-specific manner, through creating novel blue GECI (XCaMP-B) and yellow GECI (XCaMP-Y). We then defined conditions to co-express and co-measure multi-color GECIs in vivo, and succeeded in recording neuronal activity in synaptic pre- and postsynaptic pairs. Furthermore, based on the principle of the synthetic activity-responsive promoter E-SARE, we created a transgenic mouse line in which long-term memory manipulation experiments could be performed in a genetically tagged memory trace network.

Academic Significance and Societal Importance of the Research Achievements

分子生物学・生化学・解剖学・生理学・神経細胞生物学等の手法を結集して、長期記憶の想起を制御する神経細胞ネットワークを同定し、個々の細胞活動をin vivo脳で計測・操作することは、脳神経科学の大きなチャレンジのひとつである。本研究により、従来のシナプス可塑性シグナル解析と遺伝子改変動物の行動学的長期記憶解析との間の大きな溝を埋め、覚醒非拘束動物において記憶想起に必要かつ十分な細胞集団の神経活動動態計測が実現する端緒となる成果が得られた。

Research Products

• [Journal Article] Rational engineering of XCaMPs, a multicolor GECI suite for in vivo imaging of complex brain circuit dynamics. (2019)
  • Author(s): Inoue M, Takeuchi A, Manita S, Horigane S-i, Sakamoto M, Kawakami R, Yamaguchi K, Otomo K, Yokoyama H, Kim R, Yokoyama T, Takemoto-Kimura S, Abe M, Okamura M, Kondo Y, Quirin S, Ramakrishnan C, Imamura T, Sakimura K, Nemoto T, Kano M, Fujii H, Deisseroth K, Kitamura K, Bito H.
  • Journal Title: Cell Volume: 177 Issue: 5 Pages: 1346-1360
  • DOI: 10.1016/j.cell.2019.04.007
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Involvement of SRF coactivator MKL2 in BDNF-mediated activation of the synaptic activity-responsive element in the Arc gene. (2019)
  • Author(s): Kikuchi K, Ihara D, Fukuchi M, Tanabe H, Ishibashi Y, Tsujii J, Tsuda M, Kaneda M, Sakagami H, Okuno H, Bito H, Yamazaki Y, Ishikawa M, Tabuchi A.
  • Journal Title: Journal of Neurochemistry Volume: 148 Issue: 2 Pages: 204-218
  • DOI: 10.1111/jnc.14596
  • Peer Reviewed / Open Access
• [Journal Article] Long-term consolidation of ensemble neural plasticity patterns in hippocampal area CA1. (2018)
  • Author(s): Attardo A, Lu J, Kawashima T, Okuno H, Fitzgerald JE, Bito H, Schnitzer MJ.
  • Journal Title: Cell Reports Volume: 25 Issue: 3 Pages: 640-650
  • DOI: 10.1016/j.celrep.2018.09.064
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Presentation] Multiplex imaging of neural activity and signaling dynamics. (2017)
  • Author(s): Bito H
  • Organizer: Brain Science Institute Seminar, Korean Institute of Science and Technology, Seoul, Korea.
  • Invited
• [Presentation] CREB-Arc signaling in long-term memory formation. (2017)
  • Author(s): Bito H
  • Organizer: Department of Neuroscience Seminar, Karolinska Institutet, Stockholm, Sweden.
  • Invited