Functional differences of Na+/Ca2+exchanger expression in Ca2+transport system of smooth muscle of guinea pig stomach

Abstract

The plasma membrane ATP-dependent Ca2+pump and the Na+/Ca2+exchanger (NCX) are the major means of Ca2+extrusion in smooth muscle. However, little is known regarding distribution and function of the NCX in guinea pig gastric smooth muscle. The expression pattern and distribution of NCX isoforms suggest a role as a regulator of Ca2+transport in cells. Na+pump inhibition and the consequent to removal of K+caused gradual contraction in fundus. In contrast, the response was significantly less in antrum. Western blotting analysis revealed that NCX1 and NCX2 are the predominant NCX isoforms expressed in stomach, the former was expressed strongly in antrum, whereas the latter displayed greater expression in fundus. Isolated plasma membrane fractions derived from gastric fundus smooth muscle were also investigated to clarify the relationship between NCX protein expression and function. Na+-dependent Ca2+uptake increased directly with Ca2+concentration. Ca2+uptake in Na+-loaded vesicles was markedly elevated in comparison with K+-loaded vesicles. Additionally, Ca2+uptake by the Na+- or K+-loaded vesicles was substantially higher in the presence of A23187 than in its absence. The result can be explained based on the assumption that Na+gradients facilitate downhill movement of Ca2+. Na+-dependent Ca2+uptake was abolished by the monovalent cationic ionophore, monensin. NaCl enhanced Ca2+efflux from vesicles, and this efflux was significantly inhibited by gramicidin. Results documented evidence that NCX2 isoform functionally contributes to Ca2+extrusion and maintenance of contraction-relaxation cycle in gastric fundus smooth muscle.Key words: stomach, smooth muscle, Na+/Ca2+exchanger (NCX), NCX2.