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Unité Mixte de Recherche Centre National de la Recherche Scientifique 6548, Université de Nice-Sophia Antipolis, O6108 Nice Cedex 2, France
Cl
currents induced by cell swelling were characterized in an immortalized
cell line (DC1) derived from rabbit distal bright convoluted tubule by
the whole cell patch-clamp techniques and by
125I efflux experiments. Exposure of cells to
a hypotonic shock induced outwardly rectifying Cl
currents that could be blocked by 0.1 mM
5-nitro-2-(3-phenylpropyl-amino)benzoic acid, 1 mM DIDS, and by 1 mM
diphenylamine-2-carboxylate. 125I efflux
experiments showed that exposure of the monolayer to a hypotonic medium
increased 125I loss. Preincubation of cells
with LaCl3 or GdCl3 prevented the development
of the response. The addition of 10 µM adenosine to the bath medium
activated outwardly rectifying whole cell currents similar to those
recorded after hypotonic shock. This conductance was inhibited by the
A1-receptor antagonist 8-cyclopentyl-1,3-diproxylxanthine (DPCPX), LaCl3, or GdCl3 and was activated by
GTP
S. The selective A1-receptor agonist
N6-cyclopentyladenosine (CPA) mimicked the
effect of hypotonicity on 125I efflux. The
CPA-induced increase of 125I efflux was
inhibited by DPCPX and external application of LaCl3 or
GdCl3. Adenosine also enhanced Mn2+ influx
across the apical membrane. Overall, the data show that DC1 cells
possess swelling- and adenosine-activated Cl conductances
that share identical characteristics. The activation of both
conductances involved Ca2+ entry into the cell, probably
via mechanosensitive Ca2+ channels. The effects of
adenosine are mediated via A1 receptors that could mediate
the purinergic regulation of the volume-sensitive Cl conductance.
cell volume; kidney; A1 receptor
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