pH dependence of Cl/HCO3 exchanger in the rat jejunal enterocyte.
During bicarbonate absorption in rat jejunum, a Cl/HCO3 exchanger mediates bicarbonate extrusion across the basolateral membrane of the enterocyte. Previous studies demonstrated that anion antiport exhibits a particular behaviour: its activity is positively affected by the presence of sodium, but the cation is not translocated by the carrier protein. In view of the particular features of the jejunal Cl/HCO3 antiporter, first we performed a pharmacological characterisation of the transport protein using various Cl channels blockers. Then, since it is well known that anion exchangers play a substantial role in cell pH regulation, we investigated the possible involvement of jejunal basolateral Cl/HCO3 antiporter in intracellular pH maintenance. The sensitivity of the exchanger to pH was investigated by measuring 36Cl uptake into basolateral membrane vesicles either varying simultaneously intra- and extravesicular pH, or presetting at 7.4 external pH and varying only the internal one. Experiments were performed both in the absence and in the presence of Na. In all the tested conditions, uptake peaked at pH of about 7. 3-7.4 and then decreased, suggesting that the main function of Cl/HCO3 exchanger is related to HCO3 absorption rather than to intracellular pH control. Since pH-regulating mechanisms counteracting acidification are well known in the jejunal enterocyte, we investigated how it regulates pH after alkalinisation of the cytosol. We tested both basolateral and brush border membrane vesicles for the presence of a K/H exchanger, but we could not give evidence for its presence by means of 86Rb uptake experiments. In conclusion, the jejunal enterocyte seems to lack a mechanism counteracting cellular alkalinisation: the main purpose of pH homeostasis might be to hinder acidification of the cytosol due to influx of protons and production of acid by the metabolism.
Tosco
M
M
Dipartimento di Fisiologia e Biochimica Generali, Università di Milano, via Celoria 26, I-20133 Milan, Italy. marisa.tosco@unimi.it
Orsenigo
M N
MN
Gastaldi
G
G
Faelli
A
A
D017878:4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid / Q000494:pharmacology; D000818:Animals; D000838:Anions; D017920:Antiporters / Q000032:analysis / Q000037:antagonists & inhibitors / Q000378:metabolism*; D002259:Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone / Q000494:pharmacology; D018118:Chloride Channels / Q000037:antagonists & inhibitors; D027963:Chloride-Bicarbonate Antiporters; D006863:Hydrogen-Ion Concentration; D007408:Intestinal Absorption; D007583:Jejunum / Q000378:metabolism*; D008297:Male; D008871:Microvilli / Q000378:metabolism; D027961:Potassium-Hydrogen Antiporters; D051381:Rats; D017208:Rats, Wistar; D012414:Rubidium Radioisotopes / Q000378:metabolism
D000838:Anions; D017920:Antiporters; D018118:Chloride Channels; D027963:Chloride-Bicarbonate Antiporters; D027961:Potassium-Hydrogen Antiporters; D012414:Rubidium Radioisotopes; D002259:Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone; D017878:4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid