Department of Biology and Neuroscience Program, Amherst College, Amherst, MA, USA.
Methods Mol Biol. 2021;2191:201-220. doi: 10.1007/978-1-0716-0830-2_13.
Zebrafish are an excellent model organism to study many aspects of vertebrate sensory encoding and behavior. Their escape responses begin with a C-shaped body bend followed by several swimming bouts away from the potentially threatening stimulus. This highly stereotyped motor behavior provides a model for studying startle reflexes and the neural circuitry underlying multisensory encoding and locomotion. Channelrhodopsin (ChR2) can be expressed in the lateral line and ear hair cells of zebrafish and can be excited in vivo to elicit these rapid forms of escape. Here we review our methods for studying transgenic ChR2-expressing zebrafish larvae, including screening for positive expression of ChR2 and recording field potentials and high-speed videos of optically evoked escape responses. We also highlight important features of the acquired data and provide a brief review of other zebrafish research that utilizes or has the potential to benefit from ChR2 and optogenetics.
斑马鱼是研究脊椎动物感觉编码和行为的优秀模式生物。它们的逃避反应首先是一个 C 形的身体弯曲,然后是几次远离潜在威胁刺激的游泳。这种高度刻板的运动行为为研究惊跳反射和多感觉编码及运动的神经回路提供了模型。通道视紫红质(ChR2)可在斑马鱼的侧线和耳毛细胞中表达,并可在体内被激发以引发这些快速的逃避反应。在这里,我们回顾了我们研究转基因 ChR2 表达的斑马鱼幼虫的方法,包括筛选 ChR2 的阳性表达和记录场电位以及光诱发逃避反应的高速视频。我们还强调了所获得数据的重要特征,并简要回顾了其他利用或有可能受益于 ChR2 和光遗传学的斑马鱼研究。