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Towards whole-brain activity imaging to enable comprehensive understandings of neural functions


開催日 2015/7/28
時間 8:50 - 11:00
会場 Room 5 (Room 501, Kobe International Conference Center)
Chairperson(s) 飯野 雄一 / YUICHI IINO ( 東京大学大学院理学系研究科生物科学専攻 / The University of Tokyo )
石原 健 / Takeshi Ishihara (九州大学大学院理学研究院 / Department of Biology, Faculty of Sciences, Kyushu University)
  • 1S05a-1   

Real-time visualization of neuronal activity in zebrafish and C. elegans

  • 中井 淳一 / Junichi Nakai:1 大倉 正道 / Masamichi Ohkura:1,2 永村 ゆう子 / Yuko Kagawa-Nagamura:1,2 武藤 彩 / Akira Muto:3 井上 昌俊 / Masatoshi Inoue:4 尾藤 晴彦 / Haruhiko Bito:4 川上 浩一 / Koichi Kawakami:3 安藤 恵子 / Keiko Gengyo-Ando:1,2 
  • 1:埼玉大理工研 / Grad Sch Sci Eng, Saitama Univ, Saitama Japan 2:埼玉大脳セ / Saitama Univ Brain Sci Inst, Saitama Univ, Saitama, Japan 3:遺伝研初期発生 / Div Molec Dev Biol, NIG, Mishima, Japan 4:東大医神経生化 / Dept Neurochem, Grad Sch Med, Univ of Tokyo, Tokyo, Japan 

To monitor neuronal activities calcium imaging is one of the promising methods. We have been developing genetically encoded calcium indicators (GECIs) called G-CaMPs (green GECIs) and R-CaMPs (red GECIs). Because G-CaMPs and R-CaMPs are genetically encoded and their expression can be spatially and temporally controlled, G-CaMPs and R-CaMPs are particularly useful to monitor neuron- and glial-activities in vivo. To monitor neural activity in zebrafish, we expressed G-CaMP7a in the tectum, which is the visual center in the zebrafish brain, and performed calcium imaging under a fluorescent microscope. When a prey paramecium swam around a zebrafish larva expressing G-CaMP7a in the tectum, we could detect fluorescent increases in cell bodies and dendrites. We could functionally map the tectal-neuron activities according to the position of the paramecium. Recently we developed new R-CaMP called R-CaMP2. Using the calmodulin-binding sequence of CaMKK-α and CaMKK-β in lieu of an M13 sequence resulted in high affinity and threefold-faster kinetics of Ca2+ transients than the parental probe R-CaMP1.07. We expressed R-CaMP2 in C. elegans neurons and/or the body wall muscles (BWMs) and imaged their activities with a confocal microscope. We could monitor neuronal or musclular activities from freely-moving C. elegans. Next we generated transgenic C. elegans that expressed channelrhodopsin 2 and G-CaMP6 in GABA neurons and R-CaMP2 in the BMWs. We successfully achieved dual-color monitoring of neuronal and musclular activities in response to photostimulation.


研究助成:Research funds : KAKENHI(21115504)

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