Optogenetic actuators with diverse spectral tuning, ion selectivity and kinetics are constantly being engineered providing powerful tools for controlling neural activity with subcellular resolution and millisecond precision. Achieving reliable and interpretable in vivo optogenetic manipulations requires reproducible actuator expression and calibration of photocurrents in target neurons. Here, we developed nine transgenic zebrafish lines for stable opsin expression and calibrated their efficacy in vivo. We first used high-throughput behavioural assays to compare opsin ability to elicit or silence neural activity. Next, we performed in vivo whole-cell electrophysiological recordings to quantify the amplitude and kinetics of photocurrents and test opsin ability to precisely control spiking. We observed substantial variation in efficacy, associated with differences in both opsin expression level and photocurrent characteristics, and identified conditions for optimal use of the most efficient opsins. Overall, our calibrated optogenetic toolkit will facilitate the design of controlled optogenetic circuit manipulations.

中文翻译：

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稳定的斑马鱼视蛋白线的校准光遗传学工具箱

具有各种光谱调节，离子选择性和动力学的光遗传致动器不断地被工程化，提供了强大的工具来控制亚细胞分辨率和毫秒精度的神经活动。要实现可靠且可解释的体内光遗传学操作，需要可再现的致动器表达和靶神经元中光电流的校准。在这里，我们开发了九种转基因斑马鱼品系来稳定视蛋白表达，并在体内校准了它们的功效。我们首先使用高通量行为分析来比较视蛋白引发或沉默神经活动的能力。接下来，我们进行了体内全细胞电生理记录，以量化光电流的幅度和动力学，并测试视蛋白的能力，以精确控制尖峰信号。我们观察到功效的实质性变化，与视蛋白表达水平和光电流特征的差异有关，并确定了最佳使用最有效视蛋白的条件。总体而言，我们经过校准的光遗传学工具包将简化受控光遗传学电路操作的设计。