Optogenetic manipulation of Gq-and Gi/o-coupled receptor signaling in neurons and heart muscle cells

Hagio, H.; Koyama, W.; Hosaka, S.; Song, A. D.; Narantsatsral, J.; Matsuda, K.; Sugihara, T.; Shimizu, T.; Koyanagi, M.; Terakita, A.; Hibi, M.

doi:10.1101/2022.10.25.513732

bioRxiv · 10.1101/2022.10.25.513732

Optogenetic manipulation of Gq-and Gi/o-coupled receptor signaling in neurons and heart muscle cells

Hagio, H.·Koyama, W.·Hosaka, S.·Song, A. D.·Narantsatsral, J.·Matsuda, K.·Sugihara, T.·Shimizu, T.·Koyanagi, M.·Terakita, A.·Hibi, M.

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Abstract

G protein-coupled receptors (GPCRs) transmit extracellular signals into the cell depending on the type of G protein. To analyze the functions of GPCR signaling, we developed optogenetic tools using animal G protein-coupled bistable rhodopsins that can be controlled into active and inactive states by light irradiation. We expressed Gq- and Gi/o-coupled bistable rhodopsins in hindbrain reticulospinal V2a neurons, which are involved in locomotion, or in cardiomyocytes of zebrafish. Light stimulation of the reticulospinal V2a neurons expressing Gq-coupled spider Rh1 resulted in an increase in the level of cytoplasmic Ca2+ and evoked swimming behavior. Light stimulation of cardiomyocytes expressing the Gi/o-coupled mosquito Opn3, pufferfish TMT opsin, or lamprey parapinopsin induced cardiac arrest, and the effect was suppressed by treatment with pertussis toxin or barium, suggesting that Gi/o-dependent regulation of inward-rectifier K+ channels controls cardiac function. These data indicate that these rhodopsins are useful for optogenetic control of GPCR-mediated signaling in neurons and cardiomyocytes in vivo. Impact statementAnimal G protein-coupled bistable rhodopsins can regulate Gq and Gi-mediated signaling in a light-dependent manner in neurons and cardiomyocytes, making them useful for analyzing roles of GPCR signaling in vivo.

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Hagio, H., Koyama, W., Hosaka, S., Song, A. D., Narantsatsral, J., Matsuda, K., Sugihara, T., Shimizu, T., Koyanagi, M., Terakita, A., Hibi, M.. 2022-10-27. Optogenetic manipulation of Gq-and Gi/o-coupled receptor signaling in neurons and heart muscle cells. https://doi.org/10.1101/2022.10.25.513732

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