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动脉平滑肌细胞内钙瞬变与自发性瞬时外向电流同步记录技术

李鹏云, 曾晓荣, 雷明, 刘智飞, 杨艳^*

泸州医学院心血管医学研究所，泸州 646000

摘要

本文旨在建立一种可同步观察细胞内信号分子和细胞膜离子通道变化之间相互关系的实时研究方法。联合应用激光扫描共聚焦显微镜(laser scanning confocal microscopy, LSCM)显微成像技术和全细胞穿孔膜片钳技术，在急性酶分离的小鼠脑动脉平滑肌细胞上同步记录自发性瞬时外向电流(spontaneous transient outward currents, STOCs)和细胞内的钙瞬变。在全细胞模式膜片钳记录平滑肌细胞膜钾电流的同时，LSCM 可准确记录到胞浆内出现的钙瞬变。此技术对于从分子水平揭示细胞内信号转导过程和离子通道相关疾病的机制有重要意义。

关键词： 激光扫描共聚焦显微镜; 膜片钳技术; 自发性瞬时外向电流; 钙瞬变

分类号：Q4-33

The technique of simultaneous recording calcium transients andspontaneous transient outward currents in arterial smooth muscle cells

LI Peng-Yun, ZENG Xiao-Rong, LEI Ming, LIU Zhi-Fei, YANG Yan^*

Institute of Cardiovasology, Luzhou Medical College, Luzhou 646000, China

Abstract

Laser scanning confocal microscopy (LSCM) and whole-cell perforated patch-clamp techniques were combined to studysimultaneously the changes of intracellular signal molecules and membrane currents. Intracellular calcium transients and spontaneoustransient outward currents (STOCs) were recorded simultaneously in freshly isolated mouse cerebral artery smooth muscle cells. Thecells loaded with fluo-4/AM were scanned with the confocal line-scan mode. Triggering voltage pulses derived from an EPC-10 patchclamp amplifier triggered the confocal line scan. The results showed that STOCs and intracellular calcium transients could be simultaneouslyrecorded in the same cell. This technique will be useful in studies of diseases caused by impairments of intracellular Ca2+signaling and related ionic channel activities, or vice versa.

Key words: laser scanning confocal microscopy; patch clamp technique; spontaneous transient outward current; Ca2+ transients

收稿日期：2009-12-21　　录用日期：2010-05-06

通讯作者：杨艳　　E-mail: wyangyan@yahoo.com.cn

引用本文：

李鹏云, 曾晓荣, 雷明, 刘智飞, 杨艳. 动脉平滑肌细胞内钙瞬变与自发性瞬时外向电流同步记录技术[J]. 生理学报 2010; 62 (3): 269-274.

LI Peng-Yun, ZENG Xiao-Rong, LEI Ming, LIU Zhi-Fei, YANG Yan. The technique of simultaneous recording calcium transients andspontaneous transient outward currents in arterial smooth muscle cells. Acta Physiol Sin 2010; 62 (3): 269-274 (in Chinese with English abstract).