Regulation of intracellular pH in resting and in stimulated parietal cells.

Microspectrofluorimetry of the pH-sensitive, fluorescent dye 2',7'-biscarboxyethyl-5 (6)-carboxyfluorescein (BCECF) was used to measure intracellular pH (pHi) in single parietal cells (PC) of intact rabbit gastric glands during resting and stimulated conditions. In 61% of PC, histamine plus isobutylmethylxanthine (IBMX) (both 100 microM) caused a small increase in pHi, ranging from 0.04 to 0.21 pH units (average delta pHi = 0.09 +/- 0.04 units over a 6-min period). In the other 39% of PC, pHi remained constant or decreased slightly (maximum decrease was 0.10 unit). The specific inhibitors omeprazole (50 microM, blocks H+- K+-ATPase), 4,4'-diisothiocyanodihydrostilbene-2,2'-disulfonic acid (H2DIDS; 100-200 microM, blocks Cl-HCO3 exchange) and amiloride (1 mM, blocks Na-H exchange) were added to both resting and stimulated PC. In stimulated PC, omeprazole caused pHi to decrease by 0.08 unit, but this inhibitor had no effect on pHi of resting PC. H2DIDS caused pHi to increase in stimulated PC five times faster compared with resting PC. Amiloride or Na-free solution (which should reverse the Na-H exchanger and cause cellular acidification) caused pHi to decrease 2.5 or 5 times, respectively, more slowly in stimulated PC compared with resting PC. Also, recovery from NH4-induced acidification (due primarily to Na-H exchange) was 1.8 times faster (measured at pHi 6.7) in resting vs. in stimulated PC. During histamine plus IBMX-induced stimulation, increased H secretion by the H+-K+-ATPase at the apical membrane is accompanied by an increase in activity of the Cl-HCO3 exchanger at the serosal membrane.(ABSTRACT TRUNCATED AT 250 WORDS)