Department of Pediatrics, NYU School of Medicine, New York, NY 10016, USA.
J Mol Cell Cardiol. 2012 Mar;52(3):596-607. doi: 10.1016/j.yjmcc.2011.12.012. Epub 2012 Jan 3.
Since ion channels move electrical charge during their activity, they have traditionally been studied using electrophysiological approaches. This was sometimes combined with mathematical models, for example with the description of the ionic mechanisms underlying the initiation and propagation of action potentials in the squid giant axon by Hodgkin and Huxley. The methods for studying ion channels also have strong roots in protein chemistry (limited proteolysis, the use of antibodies, etc.). The advent of the molecular cloning and the identification of genes coding for specific ion channel subunits in the late 1980s introduced a multitude of new techniques with which to study ion channels and the field has been rapidly expanding ever since (e.g. antibody development against specific peptide sequences, mutagenesis, the use of gene targeting in animal models, determination of their protein structures) and new methods are still in development. This review focuses on techniques commonly employed to examine ion channel function in an electrophysiological laboratory. The focus is on the K(ATP) channel, but many of the techniques described are also used to study other ion channels.
由于离子通道在其活动过程中移动电荷,因此传统上使用电生理学方法来研究它们。这有时与数学模型相结合,例如,霍奇金和赫克斯利用数学模型描述了鱿鱼巨大轴突中动作电位起始和传播的离子机制。研究离子通道的方法也深深植根于蛋白质化学(有限蛋白水解、抗体的使用等)。分子克隆的出现以及 20 世纪 80 年代末特定离子通道亚基编码基因的鉴定,引入了许多研究离子通道的新技术,此后该领域一直在迅速扩展(例如,针对特定肽序列的抗体开发、突变、在动物模型中使用基因靶向、确定其蛋白质结构),并且新方法仍在不断发展。这篇综述重点介绍了在电生理学实验室中常用的检查离子通道功能的技术。重点介绍了 K(ATP)通道,但描述的许多技术也用于研究其他离子通道。