Jiang Lin-Hua, Roger Sébastien
School of Biomedical Sciences, Faculty of Biological Sciences, University of Leeds, Leeds, UK.
EA4245, Transplantation, Immunology, Inflammation, Faculty of Medicine, University of Tours, Tours, France.
Methods Mol Biol. 2020;2041:261-273. doi: 10.1007/978-1-4939-9717-6_19.
P2X receptors (P2XRs) are ligand-gated ion channels gated by extracellular adenosine 5'-triphosphate (ATP) and play a critical role in mediating ATP-induced purinergic signaling in physiological and pathological processes. Heterologous expression of P2XR in human embryonic kidney 293 (HEK293) cells and measurement of P2XR-mediated currents using patch-clamp recording technique have been widely used to study the biophysical and pharmacological properties of these receptors. Combination of electrophysiology with site-directed mutagenesis and structural information has shed light on the molecular basis for receptor activation and mechanisms of actions by receptor antagonists and modulators. It is anticipated that such methodologies will continue helping us to provide more mechanistic understanding of P2XRs and to test novel receptor antagonists and allosteric modulators for therapeutical purposes. In this chapter, we describe protocols of transiently or stably expressing the P2XR in HEK293 cells and measuring P2XR-mediated currents by using whole-cell recording.
P2X受体（P2XRs）是由细胞外三磷酸腺苷（ATP）门控的配体门控离子通道，在生理和病理过程中介导ATP诱导的嘌呤能信号传导中起关键作用。P2XR在人胚肾293（HEK293）细胞中的异源表达以及使用膜片钳记录技术测量P2XR介导的电流已被广泛用于研究这些受体的生物物理和药理学特性。电生理学与定点诱变和结构信息的结合揭示了受体激活的分子基础以及受体拮抗剂和调节剂的作用机制。预计此类方法将继续帮助我们更深入地了解P2XRs，并测试新型受体拮抗剂和变构调节剂以用于治疗目的。在本章中，我们描述了在HEK293细胞中瞬时或稳定表达P2XR并使用全细胞记录测量P2XR介导电流的实验方案。
