A neuropsin-based optogenetic tool for precise control of Gq signaling  

作　　者：Ruicheng Dai Tao Yu Danwei Weng Heng Li Yuting Cui Zhaofa Wu Qingchun Guo Haiyue Zou Wenting Wu Xinwei Gao Zhongyang Qi Yuqi Ren Shu Wang Yulong Li Minmin Luo 

机构地区：[1]National Institute of Biological Sciences(NIBS),Beijing 102206,China [2]School of Life Sciences,Tsinghua University,Beijing 100084,China [3]School of Life Sciences,Peking University,Beijing 100871,China [4]Graduate School of Peking Union Medical College,Beijing 100730,China [5]Chinese Institute for Brain Research,Beijing 102206,China [6]Tsinghua Institute of Multidisciplinary Biomedical Research(TIMBR),Beijing 102206,China [7]Peking University-Tsinghua University-NIBS Joint Graduate Program,NIBS,Beijing 102206,China [8]State Key Laboratory of Membrane Biology,Peking University School of Life Sciences,Beijing 100871,China [9]PKU-McGovern Institute for Brain Research,Beijing 100871,China [10]Capital Medical University,Beijing 102206,China [11]Academy for Advanced Interdisciplinary Studies,Peking University,Beijing 100871,China

出　　处：《Science China(Life Sciences)》2022年第7期1271-1284,共14页中国科学（生命科学英文版）

基　　金：supported by Ministry of Science and Technology China Brain Initiative Grant(2021ZD0202803);the Research Unit of Medical Neurobiology at Chinese Academy of Medical Sciences(2019RU003);Beijing Municipal Government。

摘　　要：G_(q)-coupled receptors regulate numerous physiological processes by activating enzymes and inducing intracellular Ca^(2+)signals.There is a strong need for an optogenetic tool that enables powerful experimental control over G_(q) signaling.Here,we present chicken opsin 5(cOpn5)as the long sought-after,single-component optogenetic tool that mediates ultra-sensitive optical control of intracellular G_(q) signaling with high temporal and spatial resolution.Expressing cOpn5 in HEK 293T cells and primary mouse astrocytes enables blue light-triggered,G_(q)-dependent Ca^(2+) release from intracellular stores and protein kinase C activation.Strong Ca^(2+) transients were evoked by brief light pulses of merely 10 ms duration and at 3 orders lower light intensity of that for common optogenetic tools.Photostimulation of cOpn5-expressing cells at the subcellular and single-cell levels generated fast intracellular Ca^(2+)transition,thus demonstrating the high spatial precision of cOpn5 optogenetics.The cOpn5-mediated optogenetics could also be applied to activate neurons and control animal behavior in a circuit-dependent manner.cOpn5 optogenetics may find broad applications in studying the mechanisms and functional relevance of G_(q) signaling in both non-excitable cells and excitable cells in all major organ systems.

关 键 词：chicken opsin 5 ASTROCYTES PHOTOSTIMULATION Ca^(2+)imaging IP3 protein kinase C neural circuit 

分 类 号：Q3-3[生物学—遗传学]