Na+-Ca2+ exchange in sarcolemmal vesicles from bovine superior mesenteric artery

Abstract

These studies were designed to determine whether a Na+-Ca2+ exchanger is present in sarcolemmal vesicles from bovine superior mesenteric artery and, if so, to determine whether this transport system is qualitatively similar to that found in other excitable tissues. Vesicles, preferentially enriched in sarcolemma, were prepared by a Mg2+ aggregation and differential centrifugation technique. An inwardly directed Ca2+ gradient stimulated 22Na+ efflux and an outwardly directed Ca2+ gradient stimulated 22Na+ uptake. Similarly, an inwardly directed Na+ gradient stimulated 45Ca2+ efflux, and an outwardly directed Na+ gradient stimulated 45Ca2+ uptake. Ca2+ gradient-stimulated Na+ transport and Na+ gradient-stimulated Ca2+ transport were not due to voltage coupling between the two ions. Hence, a Na+-Ca2+ exchanger is present in these vesicles. The Na+ gradient-dependent component of Ca2+ uptake (Na+-Ca2+ exchange) was stimulated by rendering the vesicles electropositive inside, and Na+-Ca2+ exchange activity was inhibited by amiloride and quinidine in a dose-dependent fashion. These data demonstrate similarities between this mesenteric arterial smooth muscle Na+-Ca2+ exchanger and that found in other excitable tissues. In the absence of added Ca2+, amiloride-sensitive 22Na+ uptake in the vesicles was stimulated by an outwardly directed proton gradient, and an inwardly directed Na+ gradient stimulated proton efflux. Thus these vesicles also contain a Na+-H+ exchanger, which has been found in the sarcolemma of other vascular smooth muscle cells. When Na+ uptake was stimulated via Na+-H+ exchange, the subsequent uptake of Ca2+ via Na+-Ca2+ exchange was tripled. In conclusion, these studies unequivocally demonstrate that sarcolemmal-enriched vesicles from bovine superior mesenteric artery contain a Na+-Ca2+ exchanger.(ABSTRACT TRUNCATED AT 250 WORDS)