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Related Experiment Videos

Cl- interference with the epithelial Na+ channel ENaC.

Tanja Bachhuber1, Jens König, Thilo Voelcker

  • 1Institut für Physiologie, Universität Regensburg, Universitätsstrasse 31, D-93053 Regensburg, Germany.

The Journal of Biological Chemistry
|July 20, 2005
PubMed
Summary

Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) inhibits epithelial Na+ channels (ENaC) via intracellular chloride ions. This anion-dependent regulation involves ENaC

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Area of Science:

  • Molecular Biology
  • Ion Channel Physiology
  • Cellular Transport Mechanisms

Background:

  • The cystic fibrosis transmembrane conductance regulator (CFTR) is a chloride channel regulating epithelial sodium channels (ENaC).
  • Previous research suggested cytosolic domains of ENaC and intracellular chloride ions mediate CFTR-dependent ENaC inhibition.
  • The precise mechanisms and anion dependence of ENaC regulation by CFTR and other chloride channels remain incompletely understood.

Purpose of the Study:

  • To investigate the anion dependence of epithelial Na+ channel (ENaC) activity.
  • To elucidate the role of ENaC cytosolic domains in inhibition by CFTR and CLC-0.
  • To determine the contribution of intracellular chloride concentration to ENaC regulation.

Main Methods:

  • Coexpression of rat ENaC with human CFTR or CLC-0 in Xenopus oocytes.

Related Experiment Videos

  • Measurement of amiloride-sensitive Na+ currents using electrophysiology.
  • Manipulation of intracellular and extracellular chloride concentrations.
  • Site-directed mutagenesis of ENaC subunits and activation of phospholipase C.

    • Main Results:

    • CFTR and CLC-0 expression inhibited ENaC currents in a chloride-dependent manner, particularly at high intracellular chloride concentrations.
    • ENaC inhibition by chloride was attenuated in channels with truncated C-termini or altered N-termini, and in dimeric channels.
    • While cytosolic domains, especially the C-terminus of betaENaC, are implicated, CFTR regulation of ENaC appears complex and multifactorial.

    Conclusions:

    • Intracellular chloride concentration is a critical regulator of epithelial Na+ channel (ENaC) activity.
    • The carboxyl terminus of betaENaC plays a role in chloride-dependent ENaC inhibition.
    • CFTR-mediated regulation of ENaC is complex, involving chloride ions and potentially other mechanisms.