Cl-HCO3 exchange and Na-HCO3 symport in rabbit outer medullary collecting duct cells

Abstract

The mechanisms of basolateral HCO3- (or related base) transport were examined in the outer stripe of the outer medullary collecting duct (OMCDo). Rabbit OMCDo were perfused in vitro and cell pH (pHi) of principal cells (PC) and intercalated cells (IC) was monitored with 2',7'-bis(carboxyethyl)-5(6)-carboxyfluorescein (BCECF) using fluorescence ratio imaging microscopy. Basolateral HCO3- exit was stimulated either by bath HCO3- reduction or by NH3/NH4(+)-induced cell alkalinization. Rates of pHi decrease were twice as high in IC than in PC. In both cell types HCO3- exit was dependent on Cl(-)-coupled and Na(+)-coupled mechanisms; the former was dominant in IC, whereas the latter was dominant in PC. pHi of PC and IC significantly increased by bath Cl- removal in the absence of Na+ and decreased by bath Na+ removal in the absence of Cl- but with amiloride. 4,4'-Diisothiocyanostilbene-2,2'-disulfonic acid (DIDS) inhibited these pHi responses to a reduction of bath HCO3-, Cl-, or Na+, and an alkali loading. Our results suggest that 1) the capacity for basolateral HCO3- transport is twice as high in IC than in PC, 2) both PC and IC possess basolateral Na(+)-independent Cl(-)-HCO3- exchange and Na(+)-HCO3- cotransport, and 3) basolateral HCO3- transport is mediated mainly by Na(+)-HCO3- cotransport in PC and by Na(+)-independent Cl(-)-HCO3- exchange in IC.