Gaborit Nathalie, Le Bouter Sabrina, Szuts Viktoria, Varro Andras, Escande Denis, Nattel Stanley, Demolombe Sophie
INSERM U533, Nantes, France.
J Physiol. 2007 Jul 15;582(Pt 2):675-93. doi: 10.1113/jphysiol.2006.126714. Epub 2007 May 3.
The various cardiac regions have specific action potential properties appropriate to their electrical specialization, resulting from a specific pattern of ion-channel functional expression. The present study addressed regionally defined differential ion-channel expression in the non-diseased human heart with a genomic approach. High-throughput real-time RT-PCR was used to quantify the expression patterns of 79 ion-channel subunit transcripts and related genes in atria, ventricular epicardium and endocardium, and Purkinje fibres isolated from 15 non-diseased human donor hearts. Two-way non-directed hierarchical clustering separated atria, Purkinje fibre and ventricular compartments, but did not show specific patterns for epicardium versus endocardium, nor left- versus right-sided chambers. Genes that characterized the atria (versus ventricles) included Cx40, Kv1.5 and Kir3.1 as expected, but also Cav1.3, Cav3.1, Cav alpha2 delta2, Nav beta1, TWIK1, TASK1 and HCN4. Only Kir2.1, RyR2, phospholamban and Kv1.4 showed higher expression in the ventricles. The Purkinje fibre expression-portrait (versus ventricle) included stronger expression of Cx40, Kv4.3, Kir3.1, TWIK1, HCN4, ClC6 and CALM1, along with weaker expression of mRNA encoding Cx43, Kir2.1, KChIP2, the pumps/exchangers Na(+),K(+)-ATPase, NCX1, SERCA2, and the Ca(2+)-handling proteins RYR2 and CASQ2. Transcripts that were more strongly expressed in epicardium (versus endocardium) included Cav1.2, KChIP2, SERCA2, CALM3 and calcineurin-alpha. Nav1.5 and Nav beta1 were more strongly expressed in the endocardium. For selected genes, RT-PCR data were confirmed at the protein level. This is the first report of the global portrait of regional ion-channel subunit-gene expression in the non-diseased human heart. Our data point to significant regionally determined ion-channel expression differences, with potentially important implications for understanding regional electrophysiology, arrhythmia mechanisms, and responses to ion-channel blocking drugs. Concordance with previous functional studies suggests that regional regulation of cardiac ion-current expression may be primarily transcriptional.
不同的心脏区域具有与其电特化相适应的特定动作电位特性,这是由离子通道功能表达的特定模式所导致的。本研究采用基因组学方法,探讨了非病变人类心脏中区域特异性的离子通道差异表达。利用高通量实时逆转录聚合酶链反应(RT-PCR)对从15个非病变人类供体心脏分离出的心房、心室心外膜和心内膜以及浦肯野纤维中79种离子通道亚基转录本和相关基因的表达模式进行定量分析。双向非定向层次聚类将心房、浦肯野纤维和心室区分开来,但未显示心外膜与心内膜、左侧与右侧腔室的特异性模式。正如预期的那样,表征心房(相对于心室)的基因包括Cx40、Kv1.5和Kir3.1,此外还有Cav1.3、Cav3.1、Cavα2δ2、Navβ1、TWIK1、TASK1和HCN4。只有Kir2.1、RyR2、受磷蛋白和Kv1.4在心室中表达较高。浦肯野纤维的表达图谱(相对于心室)包括Cx40、Kv4.3、Kir3.1、TWIK1、HCN4、ClC6和CALM1的更强表达,以及编码Cx43、Kir2.1、KChIP2、泵/交换体Na(+)、K(+)-ATP酶、NCX1、SERCA2以及钙处理蛋白RYR2和CASQ2的mRNA的较弱表达。在心外膜(相对于心内膜)中表达更强的转录本包括Cav1.2、KChIP2、SERCA2、CALM3和钙调神经磷酸酶α。Nav1.5和Navβ1在心内膜中表达更强。对于选定的基因,RT-PCR数据在蛋白质水平得到了证实。这是关于非病变人类心脏中区域离子通道亚基基因表达全局图谱的首次报告。我们的数据表明存在显著的区域决定的离子通道表达差异,这对于理解区域电生理学、心律失常机制以及对离子通道阻断药物的反应可能具有重要意义。与先前功能研究的一致性表明,心脏离子电流表达的区域调节可能主要是转录性的。