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1 Departments of Physiology and Animal Science and 2 Molecular Veterinary Biosciences Graduate Program, University of Minnesota, St. Paul 55108; and 3 Department of Medicine, University of Minnesota, Minneapolis, Minnesota 55455
Whole cell perforated patch-clamp
experiments were performed with adult rat alveolar epithelial cells.
The holding potential was
60 mV, and depolarizing voltage steps
activated voltage-gated K+ (Kv) channels. The
voltage-activated currents exhibited a mean reversal potential of
32
mV. Complete activation was achieved at
10 mV. The currents exhibited
slow inactivation, with significant variability in the time course
between cells. Tail current analysis revealed cell-to-cell variability
in K+ selectivity, suggesting contributions of multiple Kv
-subunits to the whole cell current. The Kv channels also displayed
steady-state inactivation when the membrane potential was held at
depolarized voltages with a window current between
30 and 5 mV.
Analysis of RNA isolated from these cells by RT-PCR revealed the
presence of eight Kv
-subunits (Kv1.1, Kv1.3, Kv1.4, Kv2.2, Kv4.1,
Kv4.2, Kv4.3, and Kv9.3), three
-subunits (Kv
1.1, Kv
2.1, and
Kv
3.1), and two K+ channel interacting protein (KChIP)
isoforms (KChIP2 and KChIP3). Western blot analysis with available Kv
-subunit antibodies (Kv1.1, Kv1.3, Kv1.4, Kv4.2, and Kv4.3) showed
labeling of 50-kDa proteins from alveolar epithelial cells grown in
monolayer culture. Immunocytochemical analysis of cells from monolayers
showed that Kv1.1, Kv1.3, Kv1.4, Kv4.2, and Kv4.3 were localized to the
apical membrane. We conclude that expression of multiple Kv
-,
-,
and KChIP subunits explains the variability in inactivation gating and
K+ selectivity observed between cells and that Kv channels
in the apical membrane may contribute to basal K+ secretion
across the alveolar epithelium.
voltage-gated potassium channels; potassium ion secretion; oxygen-sensitive potassium channels; alveolar fluid clearance
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