Na+-H+ exchange is present in sarcolemmal vesicles from dog superior mesenteric artery.
The Na+ concentration inside vascular smooth muscle cells is an important regulator of vascular smooth muscle function, but the mechanisms that mediate Na+ influx are not known. We studied Na+ transport in a newly described vesicle preparation preferentially enriched in sarcolemma and obtained by Mg2+ aggregation and differential centrifugation of homogenized dog superior mesenteric artery. In the presence of an outwardly directed proton gradient (pHout = 7.5, pHin = 5.0), 1 mM 22Na+ uptake was stimulated over twofold relative to the absence of a pH gradient (pHin = 7.5 or 5.0). pH gradient-stimulated Na+ uptake was inhibited by 1 mM amiloride. 22Na efflux was stimulated by an inwardly directed proton gradient (pHin = 7.5, pHout = 5.7 vs. 7.5). The rate of proton efflux from acid-loaded vesicles was measured by acridine orange fluorescence and was stimulated by 100 mM Naout but not by Nain = Naout = 100 mM. H+ gradient-stimulated Na+ transport and Na+ gradient-stimulated H+ transport were not due to electrical coupling between the two cations. The pH gradient-stimulated component of Na+ transport in the final vesicles, an intermediate fraction, and microsomes were proportional to the respective enzyme marker activities for sarcolemma but not for sarcoplasmic reticulum or mitochondrial membranes. We conclude that Mg2+ aggregation and differential centrifugation of homogenized vascular smooth muscle yield a vesicle preparation preferentially enriched in sarcolemma. Furthermore, the sarcolemma of vascular smooth muscle contains an amiloride-sensitive Na+-H+ proton countertransport system.
D000818:Animals; D001693:Biological Transport, Active; D002259:Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone / Q000494:pharmacology; D002352:Carrier Proteins / Q000032:analysis*; D002458:Cell Fractionation; D004285:Dogs; D003576:Electron Transport Complex IV / Q000378:metabolism; D005260:Female; D006863:Hydrogen-Ion Concentration; D008297:Male; D008638:Mesenteric Arteries / Q000032:analysis*; D009131:Muscle, Smooth, Vascular / Q000032:analysis; D009251:NADPH-Ferrihemoprotein Reductase / Q000378:metabolism; D010042:Ouabain / Q000378:metabolism; D000644:Quaternary Ammonium Compounds / Q000494:pharmacology; D012508:Sarcolemma / Q000032:analysis*; D012964:Sodium / Q000378:metabolism; D017923:Sodium-Hydrogen Exchangers; D014634:Valinomycin / Q000494:pharmacology
D002352:Carrier Proteins; D000644:Quaternary Ammonium Compounds; D017923:Sodium-Hydrogen Exchangers; D014634:Valinomycin; D002259:Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone; D010042:Ouabain; D012964:Sodium; D009251:NADPH-Ferrihemoprotein Reductase; D003576:Electron Transport Complex IV; C027917:tetramethylammonium