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Related Experiment Videos

Antisense oligonucleotides against the alpha-subunit of ENaC decrease lung epithelial cation-channel activity.

L Jain1, X J Chen, B Malik

  • 1Department of Pediatrics, Emory University School of Medicine, Atlanta, Georgia 30322, USA. ljain@emory.edu

The American Journal of Physiology
|June 11, 1999
PubMed
Summary
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Lung epithelial cells use amiloride-sensitive sodium channels for fluid balance. Research suggests the alpha-subunit of the epithelial sodium channel (ENaC) is key, not necessarily all ENaC subunits, for this nonselective cation channel function.

Area of Science:

  • Cell Biology
  • Physiology
  • Molecular Biology

Background:

  • Amiloride-sensitive sodium transport in lung epithelia is vital for alveolar fluid homeostasis.
  • The epithelial sodium channel (ENaC) subunits (alpha, beta, gamma) are present in lung epithelia, but electrophysiological studies show a different, nonselective cation channel.

Purpose of the Study:

  • To investigate the relationship between cloned ENaC subunits and the amiloride-sensitive, nonselective cation channel in rat alveolar type II (ATII) cells.
  • To determine which ENaC subunit(s) are essential for the function of this channel.

Main Methods:

  • Single-channel recordings from apical membrane patches of rat ATII cells.
  • Antisense oligonucleotide methods to selectively inhibit the synthesis of alpha, beta, and gamma ENaC subunits.

Related Experiment Videos

  • Quantification of nonselective cation channel density after subunit inhibition.

    • Main Results:

    • A nonselective cation channel (20.6 pS conductance, Na+/K+ selectivity of 0.97) inhibited by amiloride was identified in ATII cells.
    • Antisense inhibition of alpha-subunit production significantly decreased the density of these nonselective cation channels.
    • Inhibition of beta- or gamma-subunits alone or together did not affect channel density or characteristics.

    Conclusions:

    • The alpha-subunit of ENaC, alone or with an unidentified protein, is likely the primary component of lung alveolar epithelial nonselective cation channels.
    • This finding challenges the assumption that functional lung ENaC channels solely comprise the canonical alpha, beta, and gamma subunits.