Jurkat细胞Kv1.3钾通道的基本电生理学特性及动力学特征  

Electrophysiological Characteristics and Kinetics of Kv1.3 Channels in the Jurkat Cell

作　　者：代中华[1,2] 谭晓秋[1,2] 闫莉[1,2] 程秀丽[1,2] 祝红[1,2] 郝维[1,2] 曹济民[1,2] 

机构地区：[1]中国医学科学院基础医学研究所 [2]北京协和医学院基础学院生理学和病理生理学系,100005

出　　处：《医学研究杂志》2012年第12期20-23,共4页Journal of Medical Research

基　　金：国家"973"计划基金资助项目(2011CB933500)

摘　　要：目的采用膜片钳技术记录Jurkat细胞膜的Kv1.3通道电流,分析通道的动力学特征,并体会成功记录的要领。方法选择T淋巴细胞性白血病Jurkat E6-1细胞系为研究对象,采用全细胞膜片钳记录方式记录Kv1.3电流,选择合适的刺激方案,分析通道的激活、失活和复活动力学特性。结果 Jurkat细胞的Kv1.3电流具有电压门控特性,并能被ShK(100pmol/L)选择性阻断。Kv1.3通道的激活时间常数(τact)、失活时间常数(τin)和复活时间常数(τrec)分别为τact=6.47±2.44ms,τin=622.49±93.90ms,τrec=6.141s。在胞内钙没有被螯合的条件下,Kv1.3通道电流常掺杂有中等电导钙激活钾通道KCa3.1电流。玻璃微电极的口径和电极内液是否含EGTA是能否记录到纯净Kv1.3电流的关键。结论 Jurkat细胞表达功能性Kv1.3通道,该通道呈现快速激活、缓慢失活和缓慢复活的动力学特征。记录Kv1.3通道须注意剔除KCa3.1电流的掺杂。该工作对研究T淋巴细胞Kv1.3通道的功能具有一定的借鉴意义。Objective Using the patch clamp technique, this study aimed to record the Kv1.3 current in Jurkat cells, analyze the channel kinetics and experience the essentials for successful recordings. Methods Kv1.3 currents were recorded by the whole - cell con- figaration in the T lymphocyte leukemia cell line Jurkat E6 - 1. With suitable stimulus scheme, being selected the kinetics of activation, inactivation and recovery of the Kv1.3 channel were also analyzed. Results The Kv1.3 channel in Jurkat cells exhibited voltage - gated characteristics and could be blocked by ShK （100pmol/L）. The time constants for activation （τrat） , inactivation （τin） and recovery （ τrec ） were 6.47 ± 2.44ms, 622.49 ±93.90ms and 6. 141s, respectively. In the absence of intracellular Ca2＋ chelation, the Kv1.3 cur- rent was always mixed with KCa3.1 currents. The size of mircelectrode caliber and EGTA in the pipette solution were two key points for a successful recording of pure Kv1.3 current. Conclusion Jurkat cells express functional Kv1.3 channels. This channel shows kinetics characterized with rapid activation, slow inactivation and slow recovery. Removing the KCa3.1 mixing is essential for the recording of pure Kv1.3 current. This study may provide experiences in the further study of Kv1.3 channel in T lymphocytes.

关 键 词：JURKAT细胞 电压门控钾通道 激活 失活 

分 类 号：R544.1[医药卫生—心血管疾病]