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pH-stat experiments in proximal renal tubules.

G Malnic1, A G Lopes, A C Cassola

  • 1Department of Physiology, Institute of Biomedical Science, University of Sao Paulo, Brazil.

The Journal of Membrane Biology
|November 1, 1990
PubMed
Summary

The pH-stat technique measures proton (H+) fluxes in kidney tubules. This method reveals saturation kinetics in H+ transport, suggesting a role for the Na+/H+ exchanger in renal tubules.

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Area of Science:

  • Nephrology
  • Physiology
  • Biophysics

Background:

  • The pH-stat technique is established for measuring proton (H+) fluxes in gastric mucosa and urinary bladder in vitro.
  • Constant mucosal pH is maintained during these measurements.

Purpose of the Study:

  • To adapt and apply the pH-stat technique for measuring H+ fluxes in renal tubules.
  • To investigate the kinetics and mechanisms of H+ transport in the proximal tubule.

Main Methods:

  • Perfused proximal tubules with double-barreled micropipettes and blocked luminal fluid with oil.
  • Used Sb/reference microelectrodes to measure pH and inject H+ or OH- ions.
  • Employed an electronic clamping circuit to maintain constant pH and cable analysis to calculate H+ fluxes per cm2.

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Main Results:

  • Apparent H(+)-ion conductance was measured and affected by acetazolamide, OH-, and H+ injection.
  • H+ fluxes exhibited saturation kinetics with respect to the pH gradient during both OH- and H+ injection.
  • H(+)-ion back-flux into the tubule lumen also showed saturation kinetics.

Conclusions:

  • The saturation kinetics of H+ fluxes suggest inhibition by luminal H+ and dependence on Na+ and H+ gradients.
  • These findings are consistent with the involvement of a saturable Na+/H+ exchanger in renal tubular H+ transport.
  • H+ flow in renal tubules is likely mediated by a membrane component, primarily the Na+/H+ exchanger.