Na(+)-Ca2+ antiporter activity of rat hepatocytes. Effect of adrenalectomy on Ca2+ uptake and release from plasma membrane vesicles.
The presence and mode of Na(+)-Ca2+ antiporter activity were studied in hepatocytes isolated from sham-operated or adrenalectomized rats and in inside-out plasma membrane vesicles isolated from rat liver. Decreasing extracellular Na+ (Na+o) immediately increased cytosolic free calcium (Ca2+i). The rise in Ca2+i was proportional to the reduction in Na+o and was caused by an increased calcium influx, presumably on the Na(+)-Ca2+ antiporter operating in the reverse mode. Perfusing the cells with Ca(2+)-free media stimulated Ca2+ efflux and decreased Ca2+i, an effect dependent on Na+o. This suggests an activation of the forward mode of Na(+)-Ca2+ exchange. There was little difference in these parameters between sham and adx groups. In contrast, steady-state calcium uptake by inside-out plasma membrane vesicles was inhibited 40% after adrenalectomy. The decreased calcium uptake was not caused by a deficiency in the ATP-dependent Ca2+ pump, whose Km and Vmax were unaffected by adrenalectomy, but by an Na(+)-dependent leak from the vesicles. Ca2+ efflux was proportional to the extravesicular Na+ concentration, suggesting that the calcium leak may take place on a Na(+)-Ca2+ antiporter. This Na(+)-dependent calcium efflux was significantly increased in vesicles prepared from adx rat livers. These results suggest that hepatocytes have functional Na(+)-Ca2+ antiporters that can operate in both forward and reverse modes. Under normal conditions, the Na(+)-Ca2+ antiporter apparently operates in the reverse mode as a Ca2+ influx pathway. The increase in Na(+)-dependent Ca2+ efflux evoked by adrenalectomy in plasma membrane vesicles could explain the recent results we obtained in hepatocytes isolated from adx rats, showing increased calcium influx, increased Ca2+i, increased intracellular calcium sequestration, and increased plasmalemmal calcium cycling.
Studer
R K
RK
Department of Physiology, University of Pittsburgh School of Medicine, PA 15261.
Borle
A B
AB
D000315:Adrenalectomy; D000818:Animals; D001693:Biological Transport, Active; D002118:Calcium / Q000378:metabolism; D002352:Carrier Proteins / Q000378:metabolism*; D002462:Cell Membrane / Q000378:metabolism; D066298:In Vitro Techniques; D008099:Liver / Q000378:metabolism*; D008297:Male; D008564:Membrane Potentials; D051381:Rats; D012964:Sodium / Q000378:metabolism*; D019831:Sodium-Calcium Exchanger
D002352:Carrier Proteins; D019831:Sodium-Calcium Exchanger; D012964:Sodium; D002118:Calcium