Kuo Ivana Y, Hill Caryl E
Department of Pharmacology, School of Medicine, Yale University, New Haven, CT, 06520, USA.
Department of Neuroscience, John Curtin School of Medical Research, Australian National University, Canberra, ACT, 0200, Australia.
Methods Mol Biol. 2017;1527:189-200. doi: 10.1007/978-1-4939-6625-7_15.
Patch clamp electrophysiology is a powerful tool that has been important in isolating and characterizing the ion channels that govern cellular excitability under physiological and pathophysiological conditions. The ability to enzymatically dissociate blood vessels and acutely isolate vascular smooth muscle cells has enabled the application of patch clamp electrophysiology to the identification of diverse voltage dependent ion channels that ultimately control vasoconstriction and vasodilation. Since intraluminal pressure results in depolarization of vascular smooth muscle, the channels that control the voltage dependent influx of extracellular calcium are of particular interest. This chapter describes methods for isolating smooth muscle cells from resistance vessels, and for recording, isolating, and characterizing voltage dependent calcium channel currents, using patch clamp electrophysiological and pharmacological protocols.
膜片钳电生理学是一种强大的工具，在分离和表征生理和病理生理条件下控制细胞兴奋性的离子通道方面发挥了重要作用。通过酶解法分离血管并急性分离血管平滑肌细胞的能力，使得膜片钳电生理学能够应用于识别多种最终控制血管收缩和舒张的电压依赖性离子通道。由于管腔内压力会导致血管平滑肌去极化，因此控制细胞外钙电压依赖性内流的通道特别受关注。本章介绍了使用膜片钳电生理和药理学方案从阻力血管中分离平滑肌细胞，以及记录、分离和表征电压依赖性钙通道电流的方法。
