Role of monocarboxylic acid transport in intracellular pH regulation of isolated proximal cells.
Isolated proximal cells were prepared from rabbit kidney cortex by mechanical dissociation. The intracytoplasmic pH (pHi) was measured in HCO3(-)-free media (external pH (pHe), 7.3) using the fluorescent dye 2,7-biscarboxyethyl-5,6-carboxyfluorescein (BCECF). Cells were acid-loaded by the nigericin technique. Addition of 70 mM Na+ to the cells caused a rapid pHi recovery, which was blocked by 0.5 mM amiloride. When the cells were exposed to 5 mM sodium butyrate in the presence of 1 mM amiloride, the H+ efflux was significantly increased and followed Michaelis-Menten kinetics. Increasing pHe from 6.4 to 7.6 at a constant pHi of 6.4 enhanced the butyrate activation of the H+ efflux. Increasing pHi from 6.5 to 7.2 at a constant pHe of 7.2 reduced the butyrate effect. 22Na uptake experiments in the presence of 1 mM amiloride showed that 1.5 mM butyrate increased the Na+ flux in the proximal cells (pHi 7.10). The efficiency of monocarboxylic anions in promoting a pHi recovery increased with the length of their straight chain (acetate less than propionate less than butyrate less than valerate). The data show that when the Na+/H+ antiporter is blocked, the proximal cells can regulate their pHi by a Na+-coupled absorption of butyrate followed by non-ionic diffusion of butyric acid out of the cell and probably also by OH- influx by means of the OH-/anion exchanger.
Bidet
M
M
Department of Biologie, CEN Saclay, Gif-sur-Yvette, France.
Merot
J
J
Tauc
M
M
Poujeol
P
P
D000584:Amiloride / Q000494:pharmacology; D000818:Animals; D002087:Butyrates / Q000494:pharmacology; D020148:Butyric Acid; D002264:Carboxylic Acids / Q000378:metabolism*; D005260:Female; D006863:Hydrogen-Ion Concentration; D066298:In Vitro Techniques; D007672:Kidney Cortex / Q000378:metabolism*; D007687:Kidney Tubules, Proximal / Q000187:drug effects / Q000378:metabolism*; D007700:Kinetics; D011190:Potassium Cyanide / Q000494:pharmacology; D011817:Rabbits; D012964:Sodium / Q000378:metabolism
D002087:Butyrates; D002264:Carboxylic Acids; D020148:Butyric Acid; D000584:Amiloride; D012964:Sodium; D011190:Potassium Cyanide