Sarcolemmal cation channels and exchangers modify the increase in intracellular calcium in cardiomyocytes on inhibiting Na+-K+-ATPase

Abstract

Although inhibition of the sarcolemmal (SL) Na+-K+-ATPase is known to cause an increase in the intracellular concentration of Ca2+([Ca2+]i) by stimulating the SL Na+/Ca2+exchanger (NCX), the involvement of other SL sites in inducing this increase in [Ca2+]iis not fully understood. Isolated rat cardiomyocytes were treated with or without different agents that modify Ca2+movements by affecting various SL sites and were then exposed to ouabain. Ouabain was observed to increase the basal levels of both [Ca2+]iand intracellular Na+concentration ([Na+]i) as well as to augment the KCl-induced increases in both [Ca2+]iand [Na+]iin a concentration-dependent manner. The ouabain-induced changes in [Na+]iand [Ca2+]iwere attenuated by treatment with inhibitors of SL Na+/H+exchanger and SL Na+channels. Both the ouabain-induced increase in basal [Ca2+]iand augmentation of the KCl response were markedly decreased when cardiomyocytes were exposed to 0–10 mM Na+. Inhibitors of SL NCX depressed but decreasing extracellular Na+from 105–35 mM augmented the ouabain-induced increase in basal [Ca2+]iand the KCl response. Not only was the increase in [Ca2+]iby ouabain dependent on the extracellular Ca2+concentration, but it was also attenuated by inhibitors of SL L-type Ca2+channels and store-operated Ca2+channels (SOC). Unlike the SL L-type Ca2+-channel blocker, the blockers of SL Na+channel and SL SOC, when used in combination with SL NCX inhibitor, showed additive effects in reducing the ouabain-induced increase in basal [Ca2+]i. These results support the view that in addition to SL NCX, SL L-type Ca2+channels and SL SOC may be involved in raising [Ca2+]ion inhibition of the SL Na+-K+-ATPase by ouabain. Furthermore, both SL Na+/H+exchanger and Na+channels play a critical role in the ouabain-induced Ca2+increase in cardiomyocytes.