Abstract
Gq-coupled receptors regulate numerous physiological processes by activating enzymes and inducing intracellular Ca2+ signals. There is a strong need for an optogenetic tool that enables powerful experimental control over Gq 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 Gq signaling with high temporal and spatial resolution. Expressing cOpn5 in HEK 293T cells and primary mouse astrocytes enables blue light-triggered, Gq-dependent Ca2+ release from intracellular stores and protein kinase C activation. Strong Ca2+ 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 Ca2+ 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 Gq signaling in both non-excitable cells and excitable cells in all major organ systems.
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Acknowledgements
This work was 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), and Beijing Municipal Government. We would like to thank members of Luo laboratory for critical discussion of the paper. We thank the Imaging cores at Chinese Institute for Brain Research, Beijing and National Institute of Biological Sciences, Beijing for technical assistance.
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Dai, R., Yu, T., Weng, D. et al. A neuropsin-based optogenetic tool for precise control of Gq signaling. Sci. China Life Sci. 65, 1271–1284 (2022). https://doi.org/10.1007/s11427-022-2122-0
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DOI: https://doi.org/10.1007/s11427-022-2122-0