Graduate School of Science, Nagoya University, Japan, Nagoya, Japan.
Graduate School of Bioagricultural Sciences, Nagoya University, Nagoya, Japan.
Elife. 2023 Aug 17;12:e83975. doi: 10.7554/eLife.83975.
Even though microbial photosensitive proteins have been used for optogenetics, their use should be optimized to precisely control cell and tissue functions in vivo. We exploited CCR4 and ChR, cation channelrhodopsins from algae, GC1, a guanylyl cyclase rhodopsin from a fungus, and photoactivated adenylyl cyclases (PACs) from cyanobacteria (PAC) or bacteria (PAC), to control cell functions in zebrafish. Optical activation of CCR4 and ChR in the hindbrain reticulospinal V2a neurons, which are involved in locomotion, induced swimming behavior at relatively short latencies, whereas activation of GC1 or PACs achieved it at long latencies. Activation of CCR4 and ChR in cardiomyocytes induced cardiac arrest, whereas activation of PAC gradually induced bradycardia. ChR activation led to an increase in intracellular Ca in the heart, suggesting that depolarization caused cardiac arrest. These data suggest that these optogenetic tools can be used to reveal the function and regulation of zebrafish neurons and cardiomyocytes.
尽管微生物光敏蛋白已被用于光遗传学，但为了精确控制体内细胞和组织的功能，它们的应用应进行优化。我们利用藻类来源的 CCR4 和 ChR、真菌来源的鸟苷酸环化酶视蛋白 GC1、蓝细菌来源的光激活腺苷酸环化酶（PAC）或细菌来源的光激活腺苷酸环化酶（PAC）来控制斑马鱼的细胞功能。光激活后脑延髓网状脊髓 V2a 神经元中的 CCR4 和 ChR，这些神经元参与运动，在较短的潜伏期内诱导游泳行为，而激活 GC1 或 PAC 则在较长的潜伏期内实现。CCR4 和 ChR 在心肌细胞中的激活导致心脏停搏，而 PAC 的激活则逐渐导致心动过缓。ChR 的激活导致心脏细胞内 Ca2+的增加，表明去极化导致心脏停搏。这些数据表明，这些光遗传学工具可用于揭示斑马鱼神经元和心肌细胞的功能和调节。
