Transport and regulatory properties of the apical Na-K-2Cl cotransporter of macula densa cells.

Academic Article

Abstract

  • NH+4/NH3 fluxes were used to probe apical Na-K-2Cl transport activity of macula densa (MD) cells from rabbit kidney. In the presence of 25 mM NaCl and 5 mM Ba2+, addition of 20 mM NH+4 to the lumen produced a profound intracellular acidification, and approximately 80% of the initial acidification rate was bumetanide sensitive. The NH+4-induced acidification rate was dependent on luminal Cl- and Na+ with apparent affinities of 17 +/- 4 mM (Hill number 1.45) and 1.0 +/- 0.3 mM, respectively. In the presence of saturating luminal NaCl concentration ([NaCl]L), blockade of basolateral Cl- efflux with 10 microM 5-nitro-2-(3-phenylpropylamino)benzoic acid (NPPB) reduced the NH+4-induced acidification rate by 51 +/- 6% (P > 0.01, n = 5). Under similar conditions, dibutyryl-cAMP (DBcAMP) + forskolin increased the NH+4-induced acidification rate by 27%, whereas it produced no detectable effect at low luminal NaCl concentration. Most of the observed DBcAMP + forskolin effect was probably due to the stimulation of the basolateral Cl- conductance, since, in the presence of basolateral NPPB, this activation was changed to a 17.1% and 16.6% inhibition of the NH+4-induced acidification rate observed at high or low [NaCl]L, respectively. We conclude that the cotransporter found in MD cells displays, with respect to other Na-K-2Cl cotransporters, a relatively high affinity for luminal Na+ and luminal Cl- and can be specifically inhibited by increases in intracellular Cl- and cAMP concentrations.
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    Published In

    Keywords

  • Animals, Bucladesine, Carrier Proteins, Cells, Cultured, Hydrogen-Ion Concentration, Kidney, Quaternary Ammonium Compounds, Rabbits, Sodium-Potassium-Chloride Symporters
  • Digital Object Identifier (doi)

    Author List

  • Laamarti MA; Bell PD; Lapointe JY
  • Start Page

  • F703
  • End Page

  • F709
  • Volume

  • 275
  • Issue

  • 5