Basolateral outward rectifier chloride channel in isolated crypts of mouse colon.
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(-) > NO(3)(-) > F(-)>> SO(4)(2-) approximately gluconate. In inside-out patches, the channel open probability was voltage dependent but insensitive to intracellular Ca(2+) 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.
Centre National de la Recherche Scientifique, Unité de Recherche en Physiologie Cellulaire, Université de Bretagne Occidentale, Brest, France.
O
O
Mignen
S
S
Egee
M
M
Liberge
B J
BJ
Harvey
D017878:4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid / Q000494:pharmacology; D000818:Animals; D000838:Anions / Q000493:pharmacokinetics; D000873:Anthracenes / Q000494:pharmacology; D001485:Basement Membrane / Q000737:chemistry; D001692:Biological Transport / Q000187:drug effects / Q000502:physiology; D018118:Chloride Channels / Q000032:analysis* / Q000037:antagonists & inhibitors / Q000502:physiology*; D003106:Colon / Q000737:chemistry* / Q000166:cytology; D004553:Electric Conductivity; D020895:Enterocytes / Q000737:chemistry / Q000502:physiology; D005260:Female; D019204:GTP-Binding Proteins / Q000502:physiology; D016244:Guanosine 5'-O-(3-Thiotriphosphate) / Q000494:pharmacology; D006160:Guanosine Triphosphate / Q000494:pharmacology; D007700:Kinetics; D008564:Membrane Potentials / Q000187:drug effects / Q000502:physiology; D051379:Mice; D009579:Nitrobenzoates / Q000494:pharmacology; D018408:Patch-Clamp Techniques
D000838:Anions; D000873:Anthracenes; D018118:Chloride Channels; D009579:Nitrobenzoates; D016244:Guanosine 5'-O-(3-Thiotriphosphate); C058176:5-nitro-2-(3-phenylpropylamino)benzoic acid; C010030:9-anthroic acid; D006160:Guanosine Triphosphate; D019204:GTP-Binding Proteins; D017878:4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid