Proton gradients in renal cortex brush-border membrane vesicles. Demonstration of a rheogenic proton flux with acridine orange.

Academic Article

Abstract

  • The fluorescence quenching of acridine orange has been used to study the formation and dissipation of acid interior pH gradients in brush-border membrane vesicles from rabbit renal cortex. Acidic interior pH gradients were produced by 1) outwardly directed gradients of Na+ or K+, and 2) the addition of vesicles equilibrated at pH 6.0 to 7.5 buffer. The rate of pH gradient dissipation was stimulated 6.3-fold by the replacement of tetramethylammonium gluconate by tetramethylammonium chloride. A further increase, of 2-fold, was seen upon the addition of carbonyl cyanide-m-chlorophenylhydrazine, demonstrating the existence of a Cl- conductance pathway. In the presence of valinomycin, the replacement of tetramethylammonium gluconate by K gluconate increased the rate of delta pH dissipation by 11-fold, demonstrating the existence of a conductive pathway for protons. This pathway for protons was also shown by the formation of an acidic interior space by an outwardly directed K gradient in the presence of valinomycin. The parallel conductive pathways for H+ and Cl- may dissipate pH and chloride gradients across the luminal membrane of the proximal tubule.
  • Authors

    Published In

    Author List

  • Reenstra WW; Warnock DG; Yee VJ; Forte JG
  • Start Page

  • 11663
  • End Page

  • 11666
  • Volume

  • 256
  • Issue

  • 22