Mechanisms of acid release in isolated gas gland cells of the European eel Anguilla anguilla

Abstract

Mechanisms of acid production and of acid release have been analyzed in isolated gas gland cells of the eel swimbladder using a cytosensor microphysiometer. Incubation of isolated cells with oxamic acid caused a dose-dependent decrease in the rate of proton release. At the highest oxamic acid concentration used (20 mmol/l), proton release was reduced by approximately 40%; incubation with sodium fluoride (10 mmol/l) or removal of glucose from the extracellular medium caused 60 and 80% reduction, respectively. NaCN had little effect on proton secretion. Proton release of isolated gas gland cells was largely dependent on the extracellular sodium concentration, and this sodium effect was in part inhibitable by amiloride. A 15-20% reduction in the rate of proton secretion was observed in the presence of 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid, an inhibitor of anion exchange. Inhibition of mammalian H(+)-K(+)-adenosinetriphosphatase with omeprazole had no effect, whereas bafilomycin, an inhibitor of vesicular H(+)-adenosinetriphosphatase, induced a 25% reduction in proton secretion. Ethoxzolamide, a membrane-permeable inhibitor of carbonic anhydrase, caused a 60% reduction in proton secretion (inhibition constant = 54.4 nmol/l). Prontosil-dextran, a membrane-impermeable sulfonamide, also reduced the proton release, thus indicating the presence of a membrane-bound carbonic anhydrase facing the extracellular space.