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1 L'institut du thorax, Inserm U533, Faculté de Médecine, Nantes, France
2
Laboratoire de Génomique Fonctionnelle LGF-CNRS UPR2580, Montpellier, France
3
Department of Physiology, University of Oxford, Oxford, UK
Even though sequencing of the mammalian genome has led to the discovery of a large number of ionic channel genes, identification of the molecular determinants of cellular electrical properties in different regions of the heart has been rarely obtained. We developed a high-throughput approach capable of simultaneously assessing the expression pattern of ionic channel repertoires from different regions of the mouse heart. By using large-scale real-time RT-PCR, we have profiled 71 channels and related genes in the sinoatrial node (SAN), atrioventricular node (AVN), the atria (A) and ventricles (V). Hearts from 30 adult male C57BL/6 mice were microdissected and RNA was isolated from six pools of five mice each. TaqMan data were analysed using the threshold cycle (Ct) relative quantification method. Cross-contamination of each region was checked with expression of the atrial and ventricular myosin light chains. Two-way hierarchical clustering analysis of the 71 genes successfully classified the six pools from the four distinct regions. In comparison with the A, the SAN and AVN were characterized by higher expression of Navß1, Navß3, Cav1.3, Cav3.1 and Cav
2
2, and lower expression of Kv4.2, Cx40, Cx43 and Kir3.1. In addition, the SAN was characterized by higher expression of HCN1 and HCN4, and lower expression of RYR2, Kir6.2, Cavß2 and Cav
4. The AVN was characterized by higher expression of Nav1.1, Nav1.7, Kv1.6, Kvß1, MinK and Cav7. Other gene expression profiles discriminate between the ventricular and the atrial myocardium. The present study provides the first genome-scale regional ionic channel expression profile in the mouse heart.
(Received 17 August 2004;
accepted after revision 21 October 2004;
first published online 21 October 2004)
Corresponding author S. Demolombe: L'institut du thorax, INSERM U533, Faculté de Médecine, 1 rue G. Veil, 44035 Nantes cedex, France. Email: sophie.demolombe{at}nantes.inserm.fr
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