title: Selective inhibition of Cl(-) conductance in toad skin by blockers of Cl(-) channels and transporters.
abstract: We compared the effects exerted by two classes of Cl(-) transport inhibitors on a Cl(-)-selective, passive anion transport route across the skin of Bufo viridis, the conductance (G(Cl)) of which can be activated by transepithelial voltage perturbation or high cAMP at short circuit. Inhibitors of antiporters (erythrosine, eosin) or cotransporters (furosemide) reduced voltage-activated G(Cl) with IC(50) of 6 +/- 1, 54 +/- 12, and 607 +/- 125 microM, respectively; they had no effect on the cAMP-induced G(Cl). The voltage for half-maximal activation of G(Cl) (V(50)) increased compared with controls, but effects on the maximal G(Cl) at more positive clamp potentials were small. Cl(-) channel blockers from the diphenylamino-2-carboxylic acid (DPC) family [dichloro-DPC, niflumic acid, flufenamic acid, and 5-nitro-2-(3-phenylpropylamino)benzoic acid] reduced the voltage-activated G(Cl) with IC(50) of 8.3 +/- 1.2, 10.5 +/- 0.6, 16.5 +/- 3.4, and 36.5 +/- 11.4 microM, respectively, and also inhibited the cAMP-induced G(Cl), albeit with slightly larger IC(50). V(50) was not significantly changed compared with controls; the maximal G(Cl) was strongly reduced. We conclude that the pathway for Cl(-) is composed of the conductive pore proper, which is blocked by the derivatives of DPC, and a separate, voltage-sensitive regulator, which is influenced by blockers of cotransporters or antiporters. This influence is partly overcome by increasing the clamp potential and removed by high concentrations of cAMP, which renders the pathway insensitive to voltage.
authors.affiliation: Department of Physiology, University of Munich, 80336 Munich, Germany. W.Nagel@lrz.uni-muenchen.de
authors.forename: W
authors.initials: W
authors.lastname: Nagel
authors.forename: P
authors.initials: P
authors.lastname: Somieski
authors.forename: U
authors.initials: U
authors.lastname: Katz
mesh_terms: D020533:Angiogenesis Inhibitors / Q000737:chemistry / Q000494:pharmacology; D000818:Animals; D027321:Anion Transport Proteins; D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000737:chemistry / Q000494:pharmacology; D002025:Bufonidae; D002034:Bumetanide / Q000737:chemistry / Q000494:pharmacology; D002121:Calcium Channel Blockers / Q000737:chemistry / Q000494:pharmacology; D002352:Carrier Proteins / Q000037:antagonists & inhibitors / Q000378:metabolism*; D018118:Chloride Channels / Q000037:antagonists & inhibitors / Q000378:metabolism*; D002712:Chlorides / Q000378:metabolism*; D016861:Cyclooxygenase Inhibitors / Q000737:chemistry / Q000494:pharmacology; D004232:Diuretics / Q000737:chemistry / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D004553:Electric Conductivity; D004801:Eosine Yellowish-(YS) / Q000737:chemistry / Q000494:pharmacology; D005439:Flufenamic Acid / Q000737:chemistry / Q000494:pharmacology; D005456:Fluorescent Dyes / Q000737:chemistry / Q000494:pharmacology; D005665:Furosemide / Q000737:chemistry / Q000494:pharmacology; D008528:Mefenamic Acid / Q000737:chemistry / Q000494:pharmacology; D008564:Membrane Potentials / Q000187:drug effects / Q000502:physiology; D009544:Niflumic Acid / Q000737:chemistry / Q000494:pharmacology; D009579:Nitrobenzoates / Q000737:chemistry / Q000494:pharmacology; D018408:Patch-Clamp Techniques; D012867:Skin / Q000378:metabolism*; D062367:ortho-Aminobenzoates / Q000737:chemistry / Q000494:pharmacology
chemical_list: D020533:Angiogenesis Inhibitors; D027321:Anion Transport Proteins; D000894:Anti-Inflammatory Agents, Non-Steroidal; D002121:Calcium Channel Blockers; D002352:Carrier Proteins; D018118:Chloride Channels; D002712:Chlorides; D016861:Cyclooxygenase Inhibitors; D004232:Diuretics; D005456:Fluorescent Dyes; D009579:Nitrobenzoates; D062367:ortho-Aminobenzoates; D002034:Bumetanide; D008528:Mefenamic Acid; C058176:5-nitro-2-(3-phenylpropylamino)benzoic acid; D009544:Niflumic Acid; D005439:Flufenamic Acid; D005665:Furosemide; C001209:fenamic acid; D004801:Eosine Yellowish-(YS)