Department of Physiology, College of Medicine, Chung-Ang University, Seoul 156-756, Korea.
Korean J Physiol Pharmacol. 2008 Apr;12(2):51-8. doi: 10.4196/kjpp.2008.12.2.51. Epub 2008 Apr 30.
Cardiac fibroblasts constitute one of the largest cell populations in the heart, and contribute to structural, biochemical, mechanical and electrical properties of the myocardium. Nonetheless, their cardiac functions, especially electrophysiological properties, have often been disregarded in studies. Ca(2+)-activated K(+) (K(Ca)) channels can control Ca(2+) influx as well as a number of Ca(2+)-dependent physiological processes. We, therefore, attempted to identify and characterize K(Ca) channels in rat Cardiac fibroblasts. First, we showed that the cells cultured from the rat ventricle were cardiac fibroblasts by immunostaining for discoidin domain receptor 2 (DDR-2), a specific fibroblast marker. Secondly, we detected the expression of various K(Ca) channels by reverse transcription polymerase chain reaction (RT-PCR), and found all three family members of K(Ca) channels, including large conductance K(Ca) (BK-alpha1- and -beta14 subunits), intermediate conductance K(Ca) (IK), and small conductance K(Ca) (SK14 subunits) channels. Thirdly, we recorded BK, IK, and SK channels by whole cell mode patch clamp technique using their specific blockers. Finally, we performed cell proliferation assay to evaluate the effects of the channels on cell proliferation, and found that the inhibition of IK channel increased the cell proliferation. These results showed the existence of BK, IK, and SK channels in rat ventricular fibroblasts and involvement of IK channel in cell proliferation.
心肌成纤维细胞构成心脏中最大的细胞群体之一,对心肌的结构、生化、机械和电学特性都有贡献。尽管如此,它们的心脏功能,尤其是电生理特性,在研究中经常被忽视。钙激活钾(K(Ca))通道可以控制钙内流以及许多依赖于钙的生理过程。因此,我们试图鉴定和表征大鼠心肌成纤维细胞中的 K(Ca)通道。首先,我们通过免疫染色显示来自大鼠心室的培养细胞是心肌成纤维细胞,特异性标志物为 discoidin 结构域受体 2(DDR-2)。其次,我们通过逆转录聚合酶链反应(RT-PCR)检测到各种 K(Ca)通道的表达,并发现了包括大电导 K(Ca)(BK-alpha1-和 -beta14 亚基)、中电导 K(Ca)(IK)和小电导 K(Ca)(SK14 亚基)在内的所有三种 K(Ca)通道家族成员。第三,我们使用它们的特异性阻断剂通过全细胞模式膜片钳技术记录 BK、IK 和 SK 通道。最后,我们进行了细胞增殖测定以评估通道对细胞增殖的影响,发现 IK 通道的抑制增加了细胞增殖。这些结果表明大鼠心室成纤维细胞中存在 BK、IK 和 SK 通道,并且 IK 通道参与细胞增殖。