Functional expression of the Na+/Ca2+ exchanger in the embryonic mouse heart

Abstract

The Na+/Ca2+ exchanger (NCX) is one of the earliest functional genes and is currently assumed to compensate at least in part for the rudimentary sarcoplasmic reticulum in the developing mouse heart. However, to date little is known about the functional expression of NCX during development. This prompted us to investigate the NCX current (INCX) in very early (embryonic day E8.5–E9.5 post coitum), early (E10.5–E11.5), middle (E13.5) and late (E16.5) stage mouse embryonic cardiomyocytes. For standard INCX measurements, [Ca2+]i was buffered to 150 nmol/l and voltage ramps were applied from +60 mV to −120 mV.At very early stages of development, we observed a prominent role of the INCX Ca2+ inward mode in elevating the cytosolic Ca2+ concentration ([Ca2+]i). Accordingly, a high INCX density was observed (+60 mV: 4.6±0.7 pA/pF, n=14). Likewise, we found a strong Ca2+ outward mode of INCX (−120 mV: −3.9±0.7 pA/pF, n=14). At later stages, however, INCX Ca2+ inward mode was reduced by 54±6% (n=15, p<0.0001) in ventricular and 68±10% (n=9, p<0.0006) in atrial cells. For the outward mode, a reduction by 43±10% (n=15, p<0.01) in ventricular and 62±11% (n=9, p<0.004) in atrial cardiomyocytes was observed. By contrast, NCX isoform expression and the reversal potential did not significantly change during development.Thus, NCX displays a prominent Ca2+ inward and outward mode during early embryonic heart development pointing to its important contribution to maintain [Ca2+]i homeostasis. The functional and protein expression of NCX declines during further development.