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Am J Physiol Renal Physiol 274: F139-F147, 1998;
0363-6127/98 $5.00
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Vol. 274, Issue 1, F139-F147, January 1998

HCOminus 3 absorption in rabbit outer medullary collecting duct: role of luminal carbonic anhydrase

Shuichi Tsuruoka and George J. Schwartz

Departments of Pediatrics and Medicine, University of Rochester School of Medicine, Rochester, New York 14642

Membrane-bound luminal carbonic anhydrase (CA) IV, by catalyzing the dehydration of carbonic acid into CO2 plus water, facilitates H+ secretion in the renal outer medullary collecting duct from the inner stripe (OMCDi). To examine the role of CA IV on H+ secretion, we measured net HCO-3 transport in perfused OMCDi segments and examined the effect on transport of two extracellular CA inhibitors, benzolamide and F-3500, aminobenzolamide coupled to a nontoxic polymer, polyoxyethylene bis(acetic acid) [synthesized and kindly provided by C. Conroy and T. Maren (C. W. Conroy, G. C. Wynns, and T. H. Maren. Bioorg. Chem. 24: 262-272, 1996)]. These agents would inhibit only the luminal CA enzyme. Dose titration curves for net HCO-3 flux were performed for each drug. Basal HCO-3 absorptive flux was 12 pmol · min-1 · mm-1 in control segments and significantly increased to 16 pmol · min-1 · mm-1 in segments from 3-day acid-treated animals. The concentrations of benzolamide and F-3500 that inhibited HCO-3 absorption by 50% were ~0.1 and ~5 µM, similar to the Ki for CA IV inhibition by these agents (0.2 and 4.0 µM, respectively; T. Maren, C. W. Conroy, G. C. Wynns, and D. R. Godman. J. Pharmacol. Exp. Ther. 280: 98-104, 1997). Adding exogenous CA to the inhibitor in the perfusate nearly restored basal HCO-3 transport, suggesting that cytosolic CA II was not inhibited by these impermeant inhibitors. In OMCDi segments from acidotic rabbits, the concentrations of benzolamide and F-3500 that inhibited HCO-3 absorption by 50% were 50 and 500 µM, respectively, >100 times the Ki for CA IV inhibition and for inhibition of HCO-3 transport in control tubules. Thus, in the OMCDi, doses of extracellular CA inhibitors that inhibited ~50% of CA IV activity also comparably inhibited HCO-3 transport, indicating that H+ secretion depends in part on the availability of luminal CA IV activity. Acidosis substantially decreased the sensitivity of HCO-3 transport to CA inhibition.

inner stripe of outer medulla; kidney; benzolamide; aminobenzolamide; acid-base homeostasis; carbonic anhydrase IV


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