神经损伤后小鼠外周感觉神经元中阿片诱导的 Kir3 电流的膜片钳分析。

Patch Clamp Analysis of Opioid-Induced Kir3 Currents in Mouse Peripheral Sensory Neurons Following Nerve Injury.

机构信息

Department of Experimental Anesthesiology, Charité-Universitätsmedizin Berlin, Campus Benjamin Franklin, Berlin, Germany.

Institute of Biochemistry and Biology, University of Potsdam, Potsdam, Germany.

出版信息

Methods Mol Biol. 2021;2201:127-137. doi: 10.1007/978-1-0716-0884-5_12.

DOI:10.1007/978-1-0716-0884-5_12

PMID:32975795

Abstract

Patch clamp is an electrophysiological technique that allows to analyze the activity of ion channels in neurons. In this chapter, we provide a detailed description of patch clamp protocol to measure the effect of a μ-opioid receptor agonist on the activity of G protein-coupled inwardly rectifying potassium (GIRK or Kir3) channels. This is performed in peripheral sensory neurons isolated from dorsal root ganglia (DRG) of mice without or with a chronic constriction injury (CCI) of the sciatic nerve, which models neuropathic pain. We describe the induction of the CCI , isolation and culture of DRG neurons, performance of the patch clamp recordings, and identification of opioid-responding neurons.

摘要

膜片钳是一种电生理技术，可用于分析神经元中离子通道的活性。在本章中，我们提供了详细的膜片钳实验方案，以测量μ阿片受体激动剂对 G 蛋白偶联内向整流钾 (GIRK 或 Kir3) 通道活性的影响。该实验是在从小鼠背根神经节 (DRG) 分离的外周感觉神经元中进行的，而没有或有坐骨神经慢性缩窄性损伤 (CCI)，CCI 可模拟神经病理性疼痛。我们描述了 CCI 的诱导、DRG 神经元的分离和培养、膜片钳记录的进行以及阿片反应神经元的鉴定。