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[Expression of Kir2.1, SCN5a and SCN1b channel genes in mouse cardiomyocytes with various electric properties: patch clamp combined with single cell RT-PCR study.] [Article in Chinese]

LUO Hong-Yan, LIANG Hua-Min, HU Xin-Wu, TANG Ming^*

Department of Physiology, Tongji Medical College of Huazhong University of Science and Technology, Wuhan 430030, China

Abstract

This study is to explore a new method of investigating molecular basis for electrophysiological properties of early fetal cardiomyocytes. Single embryonic cardiomyocytes of mouse early developmental heart (E10.5) were obtained by a collagenase B digestion approach. After recording spontaneous action potential using whole cell patch clamp technique, the single cell was picked by a glass micropipette, followed by a standard RT-PCR to explore the expression levels of several ion channel genes. Three phenotypes of cardiomyocytes were demonstrated with distinct properties: ventricular-like, atrial-like, and pacemaker-like action potentials. Ventricular-like and atrial-like cells were characterized with much negative maximum diastolic potential (MDP) and a higher Vmax (maximum velocity of depolarization) compared to pacemaker-like cells. MDP of ventricular-like cells was the most negative. In parallel, stronger expression of SCN5a, SCN1b and Kir2.1 were observed in ventricular-like and atrial-like cells compared to that of pacemaker-like cells, where Kir2.1 in ventricular-like cells was the most abundant. Cardiomyocytes with distinct electrophysiological properties had distinct gene expression pattern. Single cell RT-PCR combined with patch clamp technique could serve as a precise detector to analyze the molecular basis of the special electrophysiological characteristics of cardiomyocytes.

Key words: patch clamp; single cell RT-PCR; electrophysiology; molecular biology; embryonic cardiomyocyte

Received: 2011-07-13　　Accepted: 2011-09-21

Corresponding author: 唐明　　E-mail: tangming49@hotmail.com

Citing This Article：

LUO Hong-Yan, LIANG Hua-Min, HU Xin-Wu, TANG Ming. [Expression of Kir2.1, SCN5a and SCN1b channel genes in mouse cardiomyocytes with various electric properties: patch clamp combined with single cell RT-PCR study.] [Article in Chinese]. Acta Physiol Sin 2012; 64 (1): 82-86 (in Chinese with English abstract).