Na-Ca exchange in ferret red blood cells

Abstract

Ferrets have high Na (140 mmol/l) red blood cells. To determine whether ferret red cells had a Na-Ca exchange system, Na effluxes via the Na + K + 2Cl cotransrpoter and Ca effluxes via the Ca pump had to be inhibited. This was accomplished by replacing cell chloride with nitrate and by loading the cells with vanadate that inhibits the Ca pump. Under these conditions, extracellular Na (Naout) inhibited Ca influx. Intracellular Na (Nain) was required for the large Ca influx as replacement of Na with Li reduced Ca influx to less than one-tenth of the original rate. Caout stimulated Na efflux by about twofold. The Ca efflux from cells depleted of Na was increased from 0.8 to 3.2 mmol.l packed cells-1.h-1 by the presence of Naout. Cells placed in a Na-free solution accumulated Ca: total intracellular Ca was 20-fold higher than free Caout. Most of this Ca was released on addition of the Ca ionophore, A23187. Because the Na gradient had driven net Ca uphill, the fluxes of Na and Ca are coupled. In a Na-free solution, the K1/2 for Ca influx was usually approximately 10 microM (occasionally approximately 100 microM), and the maximal velocity (Vmax was 1.5-4.4 mmol.l packed cells-1.h-1. At Naout = 150 mM, K1/2 increased 5- to 150-fold. In some cells, Naout decreased Vmax by approximately fourfold, suggesting that Naout does not always compete with Caout.(ABSTRACT TRUNCATED AT 250 WORDS)