Modification of intracellular calcium concentration in cardiomyocytes by inhibition of sarcolemmal Na+/H+exchanger

Abstract

Although the Na+/H+exchanger (NHE) is considered to be involved in regulation of intracellular Ca2+concentration ([Ca2+]i) through the Na+/Ca2+exchanger, the exact mechanisms of its participation in Ca2+handling by cardiomyocytes are not fully understood. Isolated rat cardiomyocytes were treated with or without agents that are known to modify Ca2+movements in cardiomyocytes and exposed to an NHE inhibitor, 5-( N-methyl- N-isobutyl)amiloride (MIA). [Ca2+]iin cardiomyocytes was measured spectrofluorometrically with fura 2-AM in the absence or presence of KCl, a depolarizing agent. MIA increased basal [Ca2+]iand augmented the KCl-induced increase in [Ca2+]iin a concentration-dependent manner. The MIA-induced increase in basal [Ca2+]iwas unaffected by extracellular Ca2+, antagonists of the sarcolemmal (SL) L-type Ca2+channel, and inhibitors of the SL Na+/Ca2+exchanger, SL Ca2+pump ATPase and mitochondrial Ca2+uptake. However, the MIA-induced increase in basal [Ca2+]iwas attenuated by inhibitors of SL Na+-K+-ATPase and sarcoplasmic reticulum (SR) Ca2+transport. On the other hand, the MIA-mediated augmentation of the KCl response was dependent on extracellular Ca2+concentration and attenuated by agents that inhibit SL L-type Ca2+channels, the SL Na+/Ca2+exchanger, SL Na+-K+-ATPase, and SR Ca2+release channels and the SR Ca2+pump. However, the effect of MIA on the KCl-induced increase in [Ca2+]iremained unaffected by treatment with inhibitors of SL Ca2+pump ATPase and mitochondrial Ca2+uptake. MIA and a decrease in extracellular pH lowered intracellular pH and increased basal [Ca2+]i, whereas a decrease in extracellular pH, in contrast to MIA, depressed the KCl-induced increase in [Ca2+]iin cardiomyocytes. These results suggest that NHE may be involved in regulation of [Ca2+]iand that MIA-induced increases in basal [Ca2+]i, as well as augmentation of the KCl-induced increase in [Ca2+]i, in cardiomyocytes are regulated differentially.