Induced overexpression of Na+/Ca2+ exchanger transgene: altered myocyte contractility, [Ca2+]i transients, SR Ca2+ contents, and action potential duration

Wang J, Chan TO, Zhang XQ, Gao E, Song J, Koch WJ, Feldman AM, Cheung JY. Induced overexpression of Na+/Ca2+ exchanger transgene: altered myocyte contractility, [Ca2+](i) transients, SR Ca2+ contents, and action potential duration. Am J Physiol Heart Circ Physiol 297: H590-H601, 2009. First published June 12, 2009; doi:10.1152/ajpheart.00190.2009.-We have produced mice in which expression of the rat cardiac Na+/Ca2+ exchanger (NCX1) transgene was switched on when doxycycline was removed from the feed at 5 wk. At 8 to 10 wk, NCX1 expression in induced (Ind) mouse hearts was 2.5-fold higher but protein levels of sarco(endo) plasmic reticulum Ca2+-ATPase, alpha(1)-and alpha(2)-subunits of Na+-K+-ATPase, phospholamban, ryanodine receptor, calsequestrin, and unphosphorylated and phosphorylated phospholemman were unchanged compared with wild-type (WT) or noninduced (non-Ind) hearts. There was no cellular hypertrophy since WT, non-Ind, and Ind myocytes had similar whole cell membrane capacitance. In Ind myocytes, NCX1 current amplitude was similar to 42% higher, L-type Ca2+ current amplitude was unchanged, and action potential duration was prolonged compared with WT or non-Ind myocytes. Contraction and intracellular Ca2+ concentration ([Ca2+](i)) transient amplitudes in Ind myocytes were lower at 0.6, not different at 1.8, and higher at 5.0 mM extracellular Ca2+ concentration ([Ca2+](o)) compared with WT or non-Ind myocytes. Despite similar Ca2+ current amplitude and sarcoplasmic reticulum (SR) Ca2+ uptake, SR Ca2+ content at 5.0 mM [Ca2+](o) was significantly higher in Ind compared with non-Ind myocytes, indicating that NCX1 directly contributed to SR Ca2+ loading. Echocardiography demonstrated that heart rate, left ventricular mass, ejection fraction, stroke volume, and cardiac output were similar among the three groups of animals. In vivo close-chest catheterization demonstrated similar contractility and relaxation among the three groups of mice, both at baseline and after stimulation with isoproterenol. We conclude that induced expression of NCX1 transgene resulted in altered [Ca2+](i) homeostasis, myocyte contractility, and action potential morphology. In addition, heart failure did not occur 3 to 5 wk after NCX1 transgene was induced to be expressed at levels found in diseased hearts.