Abstract

The Na+/Ca2+ exchanger participates in Ca2+ homeostasis in a variety of cells and has a key role in cardiac muscle physiology. We studied in this work the exchanger of amphibian skeletal muscle, using both isolated inside-out transverse tubule vesicles and single muscle fibers. In vesicles, increasing extravesicular (intracellular) Na+ concentration cooperatively stimulated Ca2+ efflux (reverse mode), with the Hill number equal to 2.8. In contrast to the stimulation of the cardiac exchanger, increasing extravesicular (cytoplasmic) Ca2+ concentration ([Ca2+]) inhibited this reverse activity with an IC50 of 91 nM. Exchanger-mediated currents were measured at 15 degrees C in single fibers voltage clamped at -90 mV. Photolysis of a cytoplasmic caged Ca2+ compound activated an inward current (forward mode) of 23 +/- 10 nA (n = 3), with an average current density of 0.6 mu A/mu F. External Na+ withdrawal generated an outward current (reverse mode) with an average current density of 0.36 +/- 0.17 mu A/mu F (n = 6) but produced a minimal increase in cytosolic [Ca2+]. These results suggest that, in skeletal muscle, the main function of the exchanger is to remove Ca2+ from the cells after stimulation.