PHi and HCO3- dependence of proton extrusion and Cl(-)-base exchange rates in isolated rabbit parietal cells.

Abstract

In many cell types, the regulation of intracellular pH (pHi) is different in the presence vs. absence of HCO3-. We investigated the pHi and HCO3- dependence of proton extrusion and anion exchange rates in isolated rabbit parietal cells loaded with the pH-sensitive dye 2',7'-bis(carboxyethyl)-5(6)-carboxyfluorescein (BCECF). In Cl(-)-depleted parietal cells, the dependence of maximal proton efflux rate on pHi showed a strong inverse correlation but was identical in the presence and absence of HCO3-. Amiloride and Na+ removal inhibited proton extrusion rates to a similar degree with or without HCO3-, whereas 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid (DIDS) had no effect. This suggests that a Na(+)-H+ exchanger is the major acid extrusion ion transporter under these experimental conditions. In Cl(-)-containing cells, there was also some Na(+)-independent, extracellular HCO(3-)- and intracellular Cl(-)-dependent, DIDS-inhibitable pHi recovery from an acid load, most likely due to intracellular Cl(-)-extracellular HCO3- exchange. Recovery from an alkaline load was primarily mediated by anion exchange, and the dependence of maximal anion exchange rates on pHi was very different in the absence and presence of HCO3-. In its absence, maximal anion exchange (Cl(-)-OH-) rates increased slowly over the tested pHi range from 6.4 to 7.8. In the presence of HCO3-, however, there was an S-shaped dependence of maximal flux rates on pHi, with a steep increase in flux rates between 6.8 and 7.5.(ABSTRACT TRUNCATED AT 250 WORDS)