Na/Ca exchange in barnacle muscle cells has a stoichiometry of 3 Na+/1 Ca2+

Abstract

The portions of the 45Ca influx and 22Na efflux that were activated by physiological concentrations of intracellular free Ca2+, [Ca2+]i, were studied in internally perfused single giant barnacle muscle cells. Since both fluxes were activated by intracellular Ca2+ (Cai) and the Ca influx was dependent on internal Na+ (Nai), the fluxes appear to be coupled (Na/Ca exchange). Tracer Ca/Ca and Na/Na exchanges were eliminated by employing tris(hydroxymethyl)aminomethane (Tris) as the predominant external cation. Under these circumstances, the ratio of the external Ca2+ (Cao)-dependent, Cai-activated Na+ efflux to the Nai-dependent, Cai-activated Ca influx was 3.1-3.2 Na+/1 Ca2+, when the intracellular Na+ concentration, [Na+]i was either 30 or 46 mM. This is the first direct measurement of the Na/Ca exchange stoichiometry. In many types of cells, the Na/Ca exchange system appears to operate in parallel with a plasma membrane ATP-driven Ca pump that has a lower capacity (maximum velocity), but higher affinity for Ca2+ than the Na/Ca exchanger. The data on the stoichiometry and activation by internal Ca2+ imply that the turnover of the Na/Ca exchanger is modulated during periods of cell activity. When the cells are depolarized, the Na/Ca exchange system is activated by the rising [Ca2+]i, and Ca2+ entry via the exchanger is promoted. Then, at repolarization, Ca2+ exits rapidly, primarily via the exchanger. However, in resting cells, with a low [Ca2+]i, much (but not all) of the Ca2+ efflux is probably mediated by the ATP-driven Ca pump.