Specific pattern of ionic channel gene expression associated with pacemaker activity in the mouse heart
Specific pattern of ionic channel gene expression associated with pacemaker activity in the mouse heart
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DOI：
10.1113/jphysiol.2004.074047
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发表时间：
2005-01-01
影响因子：
5.5
通讯作者：
Demolombe, S
中科院分区：
文献类型：
--
作者：
Marionneau, C;Couette, B;Demolombe, S
尽管哺乳动物基因组测序发现了大量离子通道基因，但对心脏不同区域细胞电特性的分子决定因素的鉴定却很少。我们开发了一种高通量方法，能够同时评估小鼠心脏不同区域的离子通道库的表达模式。通过使用大规模实时 RT-PCR，我们分析了窦房结 (SAN)、房室结 (AVN)、心房 (A) 和心室 (V) 中的 71 个通道和相关基因。对 30 只成年雄性 C57BL/6 小鼠的心脏进行显微解剖，并从 6 个组（每组 5 只小鼠）中分离出 RNA。使用阈值循环 C(t) 相对定量方法分析 TaqMan 数据。通过心房和心室肌球蛋白轻链的表达检查每个区域的交叉污染。对 71 个基因的双向层次聚类分析成功地将来自四个不同区域的六个池分类。与A相比，SAN和AVN的特点是Navbeta1、Navbeta3、Cav1.3、Cav3.1和Cavalpha2delta2的表达较高，而Kv4.2、Cx40、Cx43和Kir3.1的表达较低。此外，SAN的特点是HCN1和HCN4表达较高，而RYR2、Kir6.2、Cavbeta2和Cavgamma4表达较低。 AVN 的特点是 Nav1.1、Nav1.7、Kv1.6、Kvbeta1、MinK 和 Cavgamma7 的较高表达。其他基因表达谱可区分心室心肌和心房心肌。本研究提供了小鼠心脏中第一个基因组规模的区域离子通道表达谱。
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 C(t) 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 Navbeta1, Navbeta3, Cav1.3, Cav3.1 and Cavalpha2delta2, 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, Cavbeta2 and Cavgamma4. The AVN was characterized by higher expression of Nav1.1, Nav1.7, Kv1.6, Kvbeta1, MinK and Cavgamma7. 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.