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应用激光捕获显微切割和实时定量 PCR 技术检测大鼠心脏传导系统中关键离子通道的表达。

Expression of key ion channels in the rat cardiac conduction system by laser capture microdissection and quantitative real-time PCR.

机构信息

Department of Cardiology, The Second Hospital of Xi'an Jiaotong University, Xi'an, Shaanxi, China.

出版信息

Exp Physiol. 2010 Sep;95(9):938-45. doi: 10.1113/expphysiol.2009.051300. Epub 2010 May 28.

Abstract

The objective of this study was to investigate the molecular basis of the inferior nodal extension (INE) in the atrioventricular junctional area that accounts for arrhythmias. The INE was separated from the adult rat heart by laser capture microdissection. The mRNA expression of ion channels was detected by quantitative real-time PCR. Hierarchical clustering was used to demonstrate clustering of expression of genes in sections. The mRNA expression of HCN4, Ca(v)3.1 and Ca(v)3.2 was high in the INE, atrioventricular node and sino-atrial node, and that of Ca(v)3.2 high in Purkinje fibres. Although the expression of HCN1 and Ca(v)1.3 was low in the rat heart, it was relatively higher in the INE, atrioventricular node and sino-atrial node than in right atrial and right ventricular (working) myocytes. Both HCN2 and Ca(v)1.2 were expressed at higher levels in working myocytes than in nodal tissues and in the INE. Hierarchical clustering analysis demonstrated that the expression of the HCN and calcium channels in INE was similar to that in the slow-response automatic cells and different from that in working myocytes and Purkinje fibres. The expression of HCN and calcium channels in the INE of the adult rat heart is similar to that of slow-response automatic cells and provides a substrate for automatic phase 4 depolarization in cells.

摘要

本研究旨在探讨房室结区下位节段(INE)的分子基础,该节段与心律失常有关。通过激光捕获显微切割将 INE 与成年大鼠心脏分离。通过定量实时 PCR 检测离子通道的 mRNA 表达。层次聚类用于展示各节段基因表达的聚类情况。HCN4、Ca(v)3.1 和 Ca(v)3.2 的 mRNA 表达在 INE、房室结和窦房结中较高,而 Ca(v)3.2 在浦肯野纤维中较高。尽管 HCN1 和 Ca(v)1.3 在大鼠心脏中的表达水平较低,但在 INE、房室结和窦房结中的表达相对高于右心房和右心室(工作)心肌细胞。HCN2 和 Ca(v)1.2 在工作心肌细胞中的表达水平均高于结组织和 INE。层次聚类分析表明,INE 中 HCN 和钙通道的表达与慢反应自动细胞相似,与工作心肌细胞和浦肯野纤维不同。成年大鼠心脏 INE 中 HCN 和钙通道的表达与慢反应自动细胞相似,为细胞的自动 4 期去极化提供了底物。

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