Kyweriga Michael, Mohajerani Majid H
Department of Neuroscience, Canadian Centre for Behavioural Neuroscience, University of Lethbridge at Lethbridge, 4401 University Dr W., Lethbridge, AB, Canada, T1K 3M4.
Methods Mol Biol. 2016;1408:251-65. doi: 10.1007/978-1-4939-3512-3_17.
Recent advances in identifying genetically unique neuronal proteins has revolutionized the study of brain circuitry. Researchers are now able to insert specific light-sensitive proteins (opsins) into a wide range of specific cell types via viral injections or by breeding transgenic mice. These opsins enable the activation, inhibition, or modulation of neuronal activity with millisecond control within distinct brain regions defined by genetic markers. Here we present a useful guide to implement this technique into any lab. We first review the materials needed and practical considerations and provide in-depth instructions for acute surgeries in mice. We conclude with all-optical mapping techniques for simultaneous recording and manipulation of population activity of many neurons in vivo by combining arbitrary point optogenetic stimulation and regional voltage-sensitive dye imaging. It is our intent to make these methods available to anyone wishing to use them.
在识别具有基因独特性的神经元蛋白方面的最新进展彻底改变了脑回路的研究。研究人员现在能够通过病毒注射或培育转基因小鼠，将特定的光敏感蛋白（视蛋白）插入到广泛的特定细胞类型中。这些视蛋白能够在由基因标记定义的不同脑区中，以毫秒级控制激活、抑制或调节神经元活动。在此，我们提供一份将该技术应用于任何实验室的实用指南。我们首先回顾所需材料和实际注意事项，并提供小鼠急性手术的详细操作说明。我们最后介绍全光学映射技术，该技术通过结合任意点光遗传学刺激和区域电压敏感染料成像，在体内同时记录和操纵许多神经元的群体活动。我们的目的是让任何希望使用这些方法的人都能获取它们。
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