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Related Experiment Videos

Urinary proteases degrade epithelial sodium channels.

S A Lewis1, C Clausen

  • 1Department of Physiology and Biophysics, University of Texas Medical Branch, Galveston 77550.

The Journal of Membrane Biology
|May 1, 1991
PubMed
Summary

Urine proteases like urokinase and plasmin irreversibly alter bladder epithelial permeability by degrading amiloride-sensitive channels. This study identifies these proteases as key factors in modifying ionic conductances in the apical membrane.

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Area of Science:

  • Physiology
  • Urology
  • Membrane Biophysics

Background:

  • Mammalian urinary bladder epithelium adapts to volume changes via vesicle insertion/withdrawal.
  • Apical and cytoplasmic vesicles possess amiloride-sensitive, cation-selective, and leak conductances.
  • Glandular kallikrein degrades epithelial sodium channels into leak conductance.

Purpose of the Study:

  • To determine if kallikrein is the sole urinary constituent altering apical membrane ionic permeability.
  • To investigate the effects of other proteases and ionic conditions on bladder epithelial permeability.

Main Methods:

  • Exposure of bladder epithelium to varying mucosal pH, urea, calcium, and osmolarity.
  • Treatment with urokinase and plasmin (serine proteases).
  • Electrophysiological analysis, including fluctuation analysis, to assess ionic conductances.

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Main Results:

  • Mucosal alterations in pH, urea, calcium, or osmolarity did not affect apical membrane conductances.
  • Urokinase and plasmin caused irreversible loss of amiloride-sensitive current and altered leak currents.
  • A third, unstable conductance appeared after protease treatment, partitioning between membrane and solution.
  • Amiloride protected amiloride-sensitive channels, but not leak pathways, from hydrolysis.

Conclusions:

  • Urokinase and plasmin, not altered ionic conditions, are responsible for irreversible modifications of apical membrane permeability.
  • Loss of amiloride-sensitive current is due to reduced sodium channel density.
  • Sequential degradation models provide estimates for single-channel currents and conductances of modified channels.