Milligan Carol J, Jiang Lin-Hua
The Florey Institute of Neuroscience and Mental Health, Melbourne Brain Centre, Parkville, VIC, Australia.
School of Biomedical Sciences, Faculty of Biological Sciences, University of Leeds, Leeds, UK.
Methods Mol Biol. 2020;2041:285-300. doi: 10.1007/978-1-4939-9717-6_21.
P2X receptors are a structurally and functionally distinctive family of ligand-gated ion channels that play important roles in mediating extracellular adenosine 5'-triphosphate (ATP) signaling in diverse physiological and pathophysiological processes. For several decades, the "manual" patch-clamp technique was regarded as the gold standard assay for investigating ion channel properties. More recently, breakthroughs in the development of automated patch-clamp technologies are enabling the study of ion channels, with much greater throughput capacities. These automated platforms, of which there are many, generate consistent, reliable, high-fidelity data. This chapter demonstrates the versatility of one of these technologies for ligand-gated ion channels, with a particular emphasis on protocols that address some of the issues of receptor desensitization that are commonly associated with P2X receptor-mediated currents.
P2X受体是一类在结构和功能上独特的配体门控离子通道家族，在多种生理和病理生理过程中介导细胞外5'-三磷酸腺苷（ATP）信号传导中发挥重要作用。几十年来，“手动”膜片钳技术一直被视为研究离子通道特性的金标准检测方法。最近，自动膜片钳技术的发展取得了突破，能够以更高的通量研究离子通道。这些自动平台有很多种，能产生一致、可靠、高保真的数据。本章展示了其中一种用于配体门控离子通道技术的多功能性，特别强调了一些解决受体脱敏问题的方案，这些问题通常与P2X受体介导的电流相关。
