Outwards currents in embryonic stem cell-derived cardiomyocytes

Abstract

The aim of the present study was to investigate the expression and functional role of outwards currents during the early stages of cardiomyogenesis. The predominant repolarizing current in early-stage, embryonic stem (ES) cell-derived cardiomyocytes was a 4-aminopyridine (4-AP) sensitive [concentration for half-maximal inhibition (IC50) 1.7 mM], transient outward current (Ito) with a current density of 10.3+/-2.1 pA/pF (n=72). We observed two additional, rapidly activating, outwardly rectifying current components, I(K),sus and Ires, in early- and late-stage cardiomyocytes. These currents were characterized by slow and no inactivation, respectively, during the depolarizing voltage step. I(K),sus was detected in about 25% of cells investigated and displayed 4-AP hypersensitivity (IC50 29 microM), whereas Ires was found in all cells of both differentiation stages and was 4-AP insensitive. In contrast to early-stage cells, Ires formed the larger portion of the aggregate, whole-cell current in late-stage, ES cell-derived cardiomyocytes. The current densities of all three current components increased during development, however, the most prominent increase was observed for I(res) from 3.6+/-0.8 pA/pF (n=72) to 8+/-1.1 pA/pF (n=35). In current-clamp recordings in early-stage, spontaneously contracting cardiomyocytes, 4-AP depolarized the cells, lengthened the action potential duration (APD) and increased the action potential frequency. In late-stage cells 4-AP had no effect on action potential frequency. We conclude that in early-stage cardiomyocytes I(to) plays an important role in controlling electrical activity.