Effect of CO2 on pHi in rabbit parietal, chief, and surface cells.
We investigated the pH recovery mechanisms in rabbit parietal, chief, and surface cells during pH shifts induced by introduction or removal of exogenous CO2-HCO3-. Intracellular pH (pHi) was measured using the fluorescent dye 2',7'-bis(2-carboxyethyl)-5(6)-carboxyfluorescin (BCECF). Gastric cells were highly purified by density gradient centrifugation and elutriation. When cells suspended in N-2-hydroxyethylpiperazene-N'-2-ethanesulfonic acid (HEPES)-100% O2, extracellular pH (pHo) 7.4, were exposed to 24 mM HCO3- -5% CO2, pHo 7.4, all cells quickly acidified by 0.3-0.4 pH units. Almost complete pH-recovery occurred within 15 min. In parietal cells, 70% of this recovery was dependent on the presence of extracellular Na+ (Nao+) and was blocked by 1 mM amiloride. The Na+-independent recovery was blocked by intracellular Cl- depletion or by 0.4 mM 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid (DIDS). In chief cells and surface cells no recovery occurred in the absence of NaO+, and 1 mM amiloride blocked pH recovery in Na+-containing buffer. On removal of HCO3- -CO2, the cells alkalinized, and subsequent pH recovery was fast, substantially extracellular Cl- (ClO-) and DIDS inhibitable in parietal cells but slow and ClO- -independent in chief and surface cells. These results suggest that during intracellular acidification the Na+-H+ exchanger is the major pH regulator in these three gastric cell types even in the presence of HCO3-. During alkalinization the Cl- -HCO3-(OH-) exchanger is the predominant pH recovery mechanism in parietal, but not in chief and surface cells. In parietal cells, this exchanger is also involved in recovery from acidification.
Seidler
U
U
Department of Surgery, Beth Israel Hospital, Boston, Massachusetts 02215.
Carter
K
K
Ito
S
S
Silen
W
W
D000584:Amiloride / Q000494:pharmacology; D000818:Animals; D002245:Carbon Dioxide / Q000494:pharmacology*; D002712:Chlorides / Q000494:pharmacology; D005744:Gastric Acid / Q000378:metabolism*; D005753:Gastric Mucosa / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D006863:Hydrogen-Ion Concentration; D066298:In Vitro Techniques; D007700:Kinetics; D008297:Male; D010295:Parietal Cells, Gastric / Q000187:drug effects / Q000502:physiology*; D011817:Rabbits; D012016:Reference Values; D012964:Sodium / Q000494:pharmacology
D002712:Chlorides; D002245:Carbon Dioxide; D000584:Amiloride; D012964:Sodium