Howard Hughes Medical Institute, University of Pennsylvania, Philadelphia, PA, USA.
Waksman Institute of Microbiology, Rutgers University, Piscataway, NJ, USA.
Methods Mol Biol. 2021;2130:221-232. doi: 10.1007/978-1-0716-0381-9_17.
Drosophila melanogaster is a powerful model organism used to study circadian rhythms, historically for elucidating the molecular basis of the clock and, more recently, for allowing for dissection of neural circuits underlying rhythmic behavior. The fly can be used to investigate the neuronal basis of complex behaviors at single-neuron resolution. Patch clamp electrophysiology permits single-neuron recording of resting membrane potential and action potential firing in response to genetic or environmental manipulations or application of drugs and neurotransmitters. Here we describe a protocol for dissecting Drosophila brains for electrophysiology, setting up and using a patch clamp system, and analyzing firing data around the circadian day and in stimulation-response experiments to test for functional neuronal connectivity in circadian circuits.
黑腹果蝇是一种强大的模式生物,用于研究生物钟,历史上用于阐明生物钟的分子基础,最近用于解析节律行为的神经回路。果蝇可用于在单细胞分辨率下研究复杂行为的神经元基础。膜片钳电生理学允许对单个神经元进行记录,以响应遗传或环境操作或药物和神经递质的应用来记录静息膜电位和动作电位的发射。这里我们描述了一种用于果蝇大脑的电生理学分离、膜片钳系统的建立和使用以及围绕生物钟日的发射数据的分析以及在刺激反应实验中的分析,以测试生物钟回路中的功能性神经元连接。