Basolateral mechanisms of intracellular pH regulation in the colonic epithelial cell line HT29 clone 19A

The intracellular pH (pHi) of the colonic tumour cell line HT29 cl.19A was studied by microspectrofluorometry using the pH-sensitive dye BCECF. Single cells within a confluent monolayer, grown in a polarized manner on permeable supports, were examined. An amiloride-sensitive Na+/H+ exchange and a stilbene-insensitive Cl-/HCO3- exchange mechanism have been identified in the basolateral membrane. Removal of Na+ from the basolateral solution caused a decrease of pHi by 0.50 +/- 0.09 unit (n = 4). Amiloride or Na(+)-free solution at the apical side had no effect on pHi. Cl- removal at the basolateral side led to an increase of pHi by 0.20 +/- 0.03 unit (n = 4) whereas apical removal had no influence on pHi. This effect was independent of Na+ and was insensitive to 0.2 mM 4,4'-diisothiocyanatodihydrostilbene-2,2'-disulphonic acid. A basolateral Cl-/HCO3- exchanger is the most likely explanation for this observation. The Na+/H+ exchange mechanism in the basolateral membrane is an acid extruder, whereas the Cl-/HCO3- exchanger is an acid loader. Both of these mechanisms are important for the maintenance of intracellular pH in HT29 cl.19A cells.