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果蝇小叶板切向细胞的电生理记录

Electrophysiological Recordings from Lobula Plate Tangential Cells in Drosophila.

作者信息

Mauss Alex S, Borst Alexander

机构信息

Max Planck Institute of Neurobiology, Am Klopferspitz 18, 82152, Martinsried, Germany.

出版信息

Methods Mol Biol. 2016;1478:321-332. doi: 10.1007/978-1-4939-6371-3_20.

Abstract

Drosophila has emerged as an important model organism for the study of the neural basis of behavior. Its main asset is the experimental accessibility of identified neurons by genetic manipulation and physiological recordings. Drosophila therefore offers the opportunity to reach an integrative understanding of the development and neural underpinnings of behavior at all processing stages, from sensing to motor control, in a single species. Here, we will provide an account of the procedures involved in recording the electrical potential of individual neurons in the visual system of adult Drosophila using the whole-cell patch-clamp method. To this end, animals are fixed to a holder and mounted below a recording chamber. The head capsule is cut open and the glial sheath covering the brain is ruptured by a combination of shearing and enzymatic digest. Neuronal somata are thus exposed and targeted by low-resistance patch electrodes. After formation of a high resistance seal, electrical access to the cell is gained by small current pulses and suction. Stable recordings of large neurons are feasible for >1 h and can be combined with controlled visual stimulation as well as genetic and pharmacological manipulation of upstream circuit elements to infer circuit function in great detail.

摘要

果蝇已成为研究行为神经基础的重要模式生物。其主要优势在于通过基因操作和生理记录可对特定神经元进行实验性研究。因此,果蝇提供了一个机会,能够在单一物种中,从感知到运动控制的所有处理阶段,全面理解行为的发育及其神经基础。在此,我们将介绍使用全细胞膜片钳方法记录成年果蝇视觉系统中单个神经元电位的相关步骤。为此,将动物固定在一个支架上,并安装在记录室下方。切开头部胶囊,通过剪切和酶消化相结合的方式使覆盖大脑的神经胶质鞘破裂。这样神经元胞体就暴露出来,并被低电阻膜片电极靶向。形成高电阻封接后,通过小电流脉冲和抽吸实现对细胞的电通路。对大型神经元进行长达1小时以上的稳定记录是可行的,并且可以结合受控的视觉刺激以及对上游电路元件的基因和药理学操作,以详细推断电路功能。

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