Differential expression of voltage-gated Ca 2+-currents in cultivated aortic myocytes

Abstract

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 ( I Ca,LVA or I Ca,HVA, respectively) based on their distinct voltage-dependences of activation and inactivation, decay kinetics using Ba 2+ 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 Ca 2+ 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.