Sodium/calcium exchange in amphibian skeletal muscle fibers and isolated transverse tubules

Abstract

The Na(+)/Ca(2+) exchanger participates in Ca(2+) 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 Ca(2+) efflux (reverse mode), with the Hill number equal to 2.8. In contrast to the stimulation of the cardiac exchanger, increasing extravesicular (cytoplasmic) Ca(2+) concentration ([Ca(2+)]) inhibited this reverse activity with an IC(50) 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 Ca(2+) compound activated an inward current (forward mode) of 23 +/- 10 nA (n = 3), with an average current density of 0.6 muA/muF. External Na(+) withdrawal generated an outward current (reverse mode) with an average current density of 0.36 +/- 0.17 muA/muF (n = 6) but produced a minimal increase in cytosolic [Ca(2+)]. These results suggest that, in skeletal muscle, the main function of the exchanger is to remove Ca(2+) from the cells after stimulation.