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.
- Author:
Hong-Yan LUO
1
;
Hua-Min LIANG
;
Xin-Wu HU
;
Ming TANG
Author Information
1. Department of Physiology, Tongji Medical College of Huazhong University of Science and Technology, Wuhan 430030, China.
- Publication Type:Journal Article
- MeSH:
Animals;
Electrophysiological Phenomena;
Female;
Fetus;
Male;
Mice;
Myocytes, Cardiac;
metabolism;
physiology;
NAV1.5 Voltage-Gated Sodium Channel;
genetics;
metabolism;
Patch-Clamp Techniques;
Potassium Channels, Inwardly Rectifying;
genetics;
metabolism;
Real-Time Polymerase Chain Reaction;
Voltage-Gated Sodium Channel beta-1 Subunit;
genetics;
metabolism
- From:
Acta Physiologica Sinica
2012;64(1):82-86
- CountryChina
- Language:Chinese
-
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 V(max) (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.
