Richard S, Neveu D, Carnac G, Bodin P, Travo P, Nargeot J
Groupe d'électrophysiologie cellulaire et moléculaire, Centre de Recherches de Biochimie Macromoléculaire, CNRS UPR 9008, INSERM U249, Montpellier, France.
Biochim Biophys Acta. 1992 Nov 10;1160(1):95-104. doi: 10.1016/0167-4838(92)90042-c.
The expression of different types of Ca(2+)-channels was studied using the whole-cell patch-clamp technique in cultured rat aortic smooth-muscle myocytes. Ca(2+)-currents were identified as either low- or high voltage-activated (ICa,LVA or ICa,HVA, respectively) based on their distinct voltage-dependences of activation and inactivation, decay kinetics using Ba2+ as the charge carrier and sensitivity to dihydropyridines. The heterogeneity in the functional expression of the two types of Ca(2+)-channels in the cultured myocytes delineated four distinct phenotypes; (i), cells exhibiting only LVA currents; (ii), cells exhibiting only HVA currents; (iii), cells exhibiting both LVA and HVA currents and (iv), cells exhibiting no current. The myocytes exclusively expressed HVA currents both during the first five days in primary culture and after the cells had reached confluence (> 15 days). In contrast, LVA currents were expressed transiently between 5 and 15 days, during which time the cells were proliferating and had transient loss of contractility. Thus, both LVA and HVA Ca(2+)-current types contribute to Ca(2+)-signalling in cultured rat aortic myocytes. However, the differential expression of the two Ca2+ current types associated with differences in contractile and proliferative phenotypes suggest that they serve distinct cellular functions. Our results are consistent with the idea that LVA current expression is important for cell proliferation.
利用全细胞膜片钳技术,在培养的大鼠主动脉平滑肌细胞中研究了不同类型钙通道的表达。根据激活和失活的不同电压依赖性、以 Ba2+ 作为电荷载体时的衰减动力学以及对二氢吡啶的敏感性,将钙电流分别鉴定为低电压激活型或高电压激活型(分别为 ICa,LVA 或 ICa,HVA)。培养的心肌细胞中两种类型钙通道功能表达的异质性描绘出四种不同的表型:(i)仅表现出 LVA 电流的细胞;(ii)仅表现出 HVA 电流的细胞;(iii)同时表现出 LVA 和 HVA 电流的细胞;(iv)不表现出电流的细胞。原代培养的前五天以及细胞达到汇合状态后(> 15 天),心肌细胞仅表达 HVA 电流。相反,LVA 电流在 5 至 15 天之间短暂表达,在此期间细胞正在增殖且收缩性暂时丧失。因此,LVA 和 HVA 两种类型的钙电流均参与培养的大鼠主动脉心肌细胞中的钙信号传导。然而,两种钙电流类型的差异表达与收缩和增殖表型的差异相关,这表明它们发挥着不同的细胞功能。我们的结果与 LVA 电流表达对细胞增殖很重要这一观点一致。