The China-America Cancer Research Institute, School of Basic Medical Sciences, Guangdong Medical University, Dongguan, P. R. China.
Department of Pharmacology and Physiology, Saint Louis University School of Medicine, St. Louis, MO, USA.
Methods Mol Biol. 2022;2510:217-237. doi: 10.1007/978-1-0716-2384-8_11.
The fundamental property of P2X7 receptor (P2X7R) channels is the transport of cations across the cell surface membrane. Electrophysiology and patch-clamp photometry are readily accessible methods of measuring this flux in a wide range of cell types. They are important tools used to characterize the functional properties of native cells studied in cell culture, in vitro tissue slices, and, in some cases, in situ single cells. Further, they are efficient methods of probing the relation of structure to function of recombinant receptors expressed in heterologous systems. Here, we provide step-by-step procedures for use of two standard recording protocols, broken-patch and perforated-patch voltage clamp. Further, we describe a third technique, called the dye-overload method, that uses simultaneous measurement of membrane current and fura-2 fluorescence to quantify the contribution of Ca flux to the ATP-gated current.
P2X7 受体 (P2X7R) 通道的基本特性是阳离子通过细胞膜表面的转运。电生理学和膜片钳光光度法是测量广泛细胞类型中这种通量的便捷方法。它们是用于描述细胞培养、体外组织切片中以及在某些情况下原位单细胞中研究的天然细胞的功能特性的重要工具。此外，它们是探测重组受体结构与异源系统表达功能关系的有效方法。在这里，我们提供了两种标准记录方案（断片和穿孔片电压钳）的逐步操作程序。此外，我们还描述了第三种技术，称为染料过载法，它使用膜电流和 fura-2 荧光的同时测量来量化 Ca 流对 ATP 门控电流的贡献。
