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Am J Physiol Gastrointest Liver Physiol 279: G277-G287, 2000;
0193-1857/00 $5.00
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Vol. 279, Issue 2, G277-G287, August 2000

Basolateral outward rectifier chloride channel in isolated crypts of mouse colon

Olivier Mignen1, Stéphane Egee1, Martine Liberge2, and Brian J. Harvey3

1 Centre National de la Recherche Scientifique, Unité de Recherche en Physiologie Cellulaire, Université de Bretagne Occidentale, 29200 Brest, France; 2 Laboratoire de Biologie et Physiologie Animales, Université des Antilles et de la Guyane, Campus Fouillote, 97159 Pointe à Pitre, Guadeloupe; and 3 Cellular Physiology Research Unit, University College Cork, Cork, Ireland

Single channel patch-clamp techniques were used to demonstrate the presence of outwardly rectifying chloride channels in the basolateral membrane of crypt cells from mouse distal colon. These channels were rarely observed in the cell-attached mode and, in the inside-out configuration, only became active after a delay and depolarizing voltage steps. Single channel conductance was 23.4 pS between -100 and -40 mV and increased to 90.2 pS between 40 and 100 mV. The channel permeability sequence for anions was: I- > SCN- > Br- > Cl- > NO3- > F- SO42- approx  gluconate. In inside-out patches, the channel open probability was voltage dependent but insensitive to intracellular Ca2+ concentration. In cell-attached mode, forskolin, histamine, carbachol, A-23187, and activators of protein kinase C all failed to activate the channel, and activity could not be evoked in inside-out patches by exposure to the purified catalytic subunit of cAMP-dependent protein kinase A. The channel was inhibited by 5-nitro-2-(3-phenylpropylamino)benzoate, 9-anthracenecarboxylic acid, and DIDS. Stimulation of G proteins with guanosine 5'-O-(3-thiotriphosphate) decreased the channel open probability and conductance, whereas subsequent addition of guanosine 5'-O-(2-thiodiphosphate) reactivated the channel.

ionic channels; colonic crypts; basolateral membrane; mouse; G protein


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