Cell pH (pHi) transients were monitored at 5-min intervals with the weak acid 5,5-[14C]dimethyloxazolidine-2,4-dione and membrane potentials were estimated from the distribution of [3H]triphenylmethylphosphonium ion in separated proximal tubules (SPT) or rabbit kidney. SPT suspensions were gassed at 37 degrees C first with 5% CO2 and then with 15% CO2. Under normal conditions, pHi rapidly fell during initial 15% CO2 acid loading and then recovered within 20 min. In the presence of 10(-3) M ouabain, which eliminated Na:H exchange as a driving force for H+ secretion, initial cell acidification was still followed by cell pH recovery, which demonstrated a sodium gradient-independent H+ extruding mechanism. In the presence of 10(-3) M ouabain plus 10(-4) M potassium cyanide, there was no pHi recovery following initial cell acidification but, on the contrary, further progressive cell acidification occurred, which is compatible with passive diffusion only of HCO-3 out of the cell. From the cyanide experiments, an apparent permeability coefficient for HCO-3 of the basolateral cell membrane was calculated; this latter result allowed the calculation of rates of passive HCO-3 diffusion and of active H+ extrusion under normal conditions and in the presence of 10(-3) M ouabain. We conclude that in the proximal tubule 1) there is a primary H+ pump additional to Na:H exchange; and 2) this primary H+ pump is responsible for about 25% of active H+ extrusion following acute CO2 cellular acid loading.

rpdb remarks: "Initial demonstration of H+ pump activity in proximal tubule" -- no entry in rpdb --