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膜片钳飞脑神经元。

Patch-Clamping Fly Brain Neurons.

机构信息

Instituto de Investigación en Biomedicina de Buenos Aires (IBioBA-CONICET), Partner Institute of the Max Planck Society, Buenos Aires C1425FQD, Argentina.

Instituto de Investigación en Biomedicina de Buenos Aires (IBioBA-CONICET), Partner Institute of the Max Planck Society, Buenos Aires C1425FQD, Argentina

出版信息

Cold Spring Harb Protoc. 2022 Aug 1;2022(8):Pdb.top107796. doi: 10.1101/pdb.top107796.

Abstract

The membrane potential of excitable cells, such as neurons and muscle cells, experiences a rich repertoire of dynamic changes mediated by an array of ligand- and voltage-gated ion channels. Central neurons, in particular, are fantastic computators of information, sensing, and integrating multiple subthreshold currents mediated by synaptic inputs and translating them into action potential patterns. Electrophysiology comprises a group of techniques that allow the direct measurement of electrical signals. There are many different electrophysiological approaches, but, because neurons are small, the whole-cell patch-clamp technique is the only applicable method for recording electrical signals from individual central neurons. Here, we provide background on patch-clamp electrophysiology in and introduce protocols for dissecting larval and adult brains, as well as for achieving whole-cell patch-clamp recordings of identified neuronal types. Patch clamping is a labor-intensive technique that requires a great deal of practice to become an expert; therefore, a steep learning curve should be anticipated. However, the instant gratification of neuronal spiking is an experience that we wish to share and disseminate, as many more patch clampers are needed to study the electrical features of so many fly neuronal types unknown to date.

摘要

可兴奋细胞(如神经元和肌肉细胞)的膜电位会经历一系列丰富的动态变化,这些变化是由一系列配体门控和电压门控离子通道介导的。特别是中枢神经元,是信息传感和整合的神奇计算器,它可以感知和整合由突触输入介导的多个亚阈电流,并将其转化为动作电位模式。电生理学是一组允许直接测量电信号的技术。有许多不同的电生理方法,但由于神经元很小,全细胞膜片钳技术是记录单个中枢神经元电信号的唯一适用方法。在这里,我们提供了膜片钳电生理学的背景知识,并介绍了分离幼虫和成年大脑的方案,以及实现对已鉴定神经元类型的全细胞膜片钳记录的方案。膜片钳是一项劳动密集型技术,需要大量的实践才能成为专家;因此,预计会有一个陡峭的学习曲线。然而,神经元放电的即时满足感是我们希望分享和传播的体验,因为需要更多的膜片钳来研究目前未知的如此多的果蝇神经元类型的电特性。

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