Proton conductance and intracellular pH recovery from an acid load in chicken enterocytes.

Abstract

1. Chicken enterocytes present a Na(+)-independent proton transport mechanism involved in pHi recovery from an acid load. In the current study the nature of this proton transport system is investigated. 2. The pHi of acid-loaded cells increased when transferred to Na(+)-free, pH 7.4 buffers, both at 6 and 65 mM extracellular potassium concentration. Addition of nigericin accelerated the rate of cell alkalinization. 3. When acid-loaded cells were transferred to a Na(+)-free, pH 6.5 buffer, the cells acidified further, regardless of the extracellular potassium concentration. The addition of nigericin increased the rate of acidification at 6 mM K+ but produced an alkalinization at 65 mM K+. 4. The rate of the Na(+)-independent regulatory cell alkalinization was inhibited by SCH 28080, DCCD, NBD-Cl, rotenone or Zn2+. Addition of valinomycin reversed the inhibition induced by SCH 28080, DCCD and NBD-Cl but not that induced by Zn2+ or rotenone. Zn2+ inhibition was abolished by the metal chelator DTPA. 5. Cytosolic acidification increased the rate of Na(+)-independent regulatory cell alkalinization. 6. The results suggest that the Na(+)-independent proton transport system is a Zn(2+)-sensitive proton-conducting pathway which is regulated by the cytosolic proton concentration.