Optogenetic manipulation of Gq- and Gi/o-coupled receptor signaling in neurons and heart muscle cells
Optogenetic manipulation of Gq- and Gi/o-coupled receptor signaling in neurons and heart muscle cells
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神经元和心肌细胞中 Gq 和 Gi/o 偶联受体信号传导的光遗传学操作
DOI：
10.1101/2022.10.25.513732
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发表时间：
2022
期刊：
影响因子：
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通讯作者：
Masahiko Hibi
中科院分区：
文献类型：
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作者：
Hanako Hagio;Wataru Koyama;Shiori Hosaka;Aysenur Deniz Song;Janchiv Narantsatsral;Koji Matsuda;Tomohiro Sugihara;Takashi Shimizu;Mitsumasa Koyanagi;Akihisa Terakita;Masahiko Hibi
G蛋白偶联受体（GPCR）根据G蛋白类型将信号传递到细胞中。为了分析GPCR信号传导的功能，我们评估了动物G蛋白偶联的视紫红质的有效性，所述视紫红质可以通过使用斑马鱼的光应用被控制成活性和非活性状态。我们表达Gq和Gi/o-耦合视紫红质在后脑网状脊髓V2 a神经元，这是参与运动，或在心肌细胞。光刺激表达Gq偶联蜘蛛Rh 1的网状脊髓V2 a神经元可引起细胞内Ca 2+水平升高并诱发游泳行为。光刺激表达Gi/o偶联的蚊子Opn 3、河豚TMT视蛋白或七鳃鳗副视蛋白的心肌细胞诱导心脏骤停，并且该作用通过用百日咳毒素或钡处理而被抑制，表明Gi/o依赖性调节内向整流K+通道控制心脏功能。这些数据表明，这些视紫红质可用于斑马鱼神经元和心肌细胞中GPCR介导的信号传导的光遗传学控制。
G-protein-coupled receptors (GPCRs) transmit signals into cells depending on the G protein type. To analyze the functions of GPCR signaling, we assessed the effectiveness of animal G-protein-coupled bistable rhodopsins that can be controlled into active and inactive states by light application using zebrafish. We expressed Gq-and Gi/o-coupled bistable rhodopsins in hindbrain reticulospinal V2a neurons, which are involved in locomotion, or in cardiomyocytes. Light stimulation of the reticulospinal V2a neurons expressing Gq-coupled spider Rh1 resulted in an increase in the intracellular Ca 2+ level 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 zebrafish neurons and cardiomyocytes.