AJP - Renal Physiology, Vol 270, Issue 1 131-F140, Copyright © 1996 by American Physiological Society

ARTICLES

Effect of cAMP agonists on cell pH and anion transport by cultured rat inner medullary collecting duct cells

C. Zhang, R. F. Husted and J. B. Stokes
Department of Internal Medicine, University of Iowa College of Medicine, Iowa City, USA.

The rat inner medullary collecting duct is capable of secreting anions. We previously showed that adenosine 3',5'-cyclic monophosphate (cAMP) stimulates anion secretion; the apical membrane anion exit pathway activated by cAMP appears to be the cystic fibrosis transmembrane conductance regulator Cl- channel. The present experiments were designed to test the hypothesis that the entry pathway across the basolateral membrane is a Cl-/HCO3- exchanger operating in parallel with an Na+/H+ exchanger. We investigated the mechanism by measuring cell Cl-, cell pH, and short-circuit current under a variety of conditions designed to uncover these pathways. cAMP agonists caused little change in cell Cl-, but they produced a consistent intracellular acidification. This acidification was dependent on HCO3-, but not on Cl-, and was not inhibited by 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid (DIDS). The presence of the basolateral Cl-/HCO3- exchanger was demonstrated by several maneuvers, and its activity was inhibited by DIDS. Applied to the basolateral solution, DIDS did not inhibit the cAMP-dependent anion current but actually stimulated it. We conclude that cAMP-stimulated anion secretion does not require activation of the basolateral Cl-/HCO3- exchanger. The transporter responsible for Cl- entry across the basolateral membrane remains unknown and is not inhibited by a variety of anion transport inhibitors, including DIDS, bumetanide, and hydrochlorothiazide. The cell acidification induced by cAMP appears to be independent of acid secretion and is the result of activation of one or more HCO3- exit pathways that are resistant to DIDS but are inhibited by a nonspecific anion transport inhibitor, 5-nitro-2-(3-phenylpro-pylamino) benzoic acid. We present a revised model for anion transport by the rat inner medullary collecting duct.

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