Generation and application of transgenic mice for wide-field and multi-color calcium imaging

• Project/Area Number: 18K19493
• Research Category: Grant-in-Aid for Challenging Research (Exploratory)
• Allocation Type: Multi-year Fund
• Review Section: Medium-sized Section 51:Brain sciences and related fields
• Research Institution: The University of Tokyo
• Principal Investigator: Sakamoto Masayuki 東京大学, 大学院医学系研究科(医学部), 助教 (00777865)
• Project Period (FY): 2018-06-29 – 2020-03-31
• Project Status: Completed (Fiscal Year 2019)
• Budget Amount: ¥6,240,000 (Direct Cost: ¥4,800,000、Indirect Cost: ¥1,440,000); Fiscal Year 2019: ¥3,120,000 (Direct Cost: ¥2,400,000、Indirect Cost: ¥720,000); Fiscal Year 2018: ¥3,120,000 (Direct Cost: ¥2,400,000、Indirect Cost: ¥720,000)
• Keywords: カルシウムイメージング / 遺伝子改変マウス / ２光子励起顕微鏡 / 多色イメージング / 遺伝的改変マウス / 2光子励起顕微鏡

Outline of Final Research Achievements

To achieve wide-field and multi-color calcium imaging with high-speed and high-sensitive calcium indicators, we generated and evaluated novel transgenic mice that express red calcium indicator and green calcium indicator in excitatory neurons inhibitory neurons, respectively. As a result, we succeeded to establish new transgenic lines that can detect fluorescent signals in response to spontaneous and sensory-evoked activity in vivo with two-photon microscopy.

Academic Significance and Societal Importance of the Research Achievements

高次脳機能を制御する神経ネットワークを理解するためには、一細胞レベルの解像度を保ちつつ、広域から多数のニューロンの活動を同時に計測する必要がある。本研究で樹立した新規遺伝子改変マウスを応用することで、興奮性ニューロンと抑制性ニューロン活動を計測することが可能となった。これらマウスを用いることで、興奮性ニューロンと抑制性ニューロンが織りなす高度に制御されたネットワークダイナミクスについて理解できるものと期待される。

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
  • DOI: 10.1016/j.cell.2019.04.007
  • URL: https://localhost/en/publications/a48843c1-543d-4414-a93c-3cab0e1df627
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Comparative Evaluation of Genetically Encoded Voltage Indicators (2019)
  • Author(s): Bando Yuki、Sakamoto Masayuki、Kim Samuel、Ayzenshtat Inbal、Yuste Rafael
  • Journal Title: Cell Reports Volume: 26 Issue: 3 Pages: 802-813
  • DOI: 10.1016/j.celrep.2018.12.088
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Presentation] Widefield sensory-evoked calcium imaging in excitatory and inhibitory neurons using a G-CaMP9a transgenic mouse. (2019)
  • Author(s): Masayuki Sakamoto, Masatoshi Inoue, Kazuki Sakai, Shigetaka Kobari, SayakaTakemoto-Kimura, Manabu Abe, Kenji Sakimura, Haruhiko Bito.
  • Organizer: Sculpted Light in the Brain
  • Int'l Joint Research
• [Presentation] 個性創発メカニズム解明のための神経活動イメージング (2019)
  • Author(s): 坂本 雅行
  • Organizer: 新学術領域「個性」創発脳 第4 回若手の会・技術支援講習会
  • Invited
• [Presentation] 膜電位感受性蛍光プローブを用いた 神経活動イメージング (2018)
  • Author(s): 坂本雅行
  • Organizer: 名古屋大学創薬科学セミナー
  • Invited
• [Presentation] Widefield sensory-evoked calcium imaging in excitatory and inhibitory neurons using a G-CaMP9a transgenic mouse (2018)
  • Author(s): Masayuki Saskamoto, Masatoshi Inoue, Kazuki Sakai, Shigetaka Kobari, SayakaTakemoto-Kimura, Manabu Abe, Kenji Sakimura, Haruhiko Bito.
  • Organizer: Japan-UK Neuroscience Symposium 2019
  • Int'l Joint Research