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Unité Mixte de Recherche 6548, Centre National de la Recherche Scientifique, Université de Nice-Sophia Antipolis, 06108 Nice Cedex 2, France
Previous
studies using the patch-clamp technique demonstrated the presence of a
small conductance Cl
channel in the apical membrane of
respiratory gill cells in primary culture originating from sea bass
Dicentrarchus labrax. We used the same technique here to
characterize potassium channels in this model. A
K+ channel of 123 ± 3 pS was identified in the
cell-attached configuration with 140 mM KCl in the bath and in the
pipette. The activity of the channel declined rapidly with time and
could be restored by the application of a negative pressure to the
pipette (suction) or by substitution of the bath solution with a
hypotonic solution (cell swelling). In the excised patch inside-out
configuration, ionic substitution demonstrated a high selectivity of
this channel for K+ over Na+ and
Ca2+. The mechanosensitivity of this channel to membrane
stretching via suction was also observed in this configuration.
Pharmacological studies demonstrated that this channel was inhibited by
barium (5 mM), quinidine (500 µM), and gadolinium (500 µM). Channel
activity decreased when cytoplasmic pH was decreased from 7.7 to 6.8. The effect of membrane distension by suction and exposure to hypotonic solutions on K+ channel activity is consistent with the
hypothesis that stretch-activated K+ channels could mediate
an increase in K+ conductance during cell swelling.
mechanosensitive K+ channels; patch clamp; gill epithelium
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