Transepithelial electrical responses to Cl- of nonsensory region of gerbil utricle.
Sheets of utricular epithelium from gerbil were mounted in a micro-Ussing chamber in order to identify and localize chloride conductances. The [Cl-] was rapidly reduced (substituted with isethionate) in the apical or basolateral perfusate and the transepithelial potential difference (Vt) and transepithelial resistance (Rt) were monitored continuously. In addition, agents known to inhibit anion transport in other epithelia were applied. The direction of all initial changes in Vt and Rt due to Cl- substitutions were consistent with the presence of ionic conductances for Cl- on both sides of the epithelium. The time-courses and magnitudes of the fall in Vt and increase in Rt during apical [Cl-] steps in the presence and absence of basolateral bumetanide were monophasic and identical in the two cases. The response of Vt to basolateral [Cl-] steps was biphasic and the initial response was greatly attenuated by bumetanide. These findings demonstrate that the largest conductance for Cl- is in the basolateral cell membrane, but that the paracellular and/or apical pathway also possess a finite Cl- conductance. All three agents tested, 3',5-dichlorodiphenylamine-2-carboxylic acid (DCDPC), 5-nitro-2(3-phenylpropylamino)benzoic acid (NPPB) and 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid (DIDS), caused an increase in Vt. NPPB and DIDS were more effective from the apical side. DCDPC and DIDS administered from the apical side led to a decrease in Rt. These results suggest that these agents act in this tissue by enhancing a conductive pathway on the apical membrane rather than blocking the basolateral Cl- conductance.
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Boys Town National Institute, Biophysics Laboratory, Omaha, NE 68131.
Marcus
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D017878:4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid; D012856:4-Acetamido-4'-isothiocyanatostilbene-2,2'-disulfonic Acid / Q000031:analogs & derivatives / Q000494:pharmacology; D000818:Animals; D002034:Bumetanide / Q000494:pharmacology; D018118:Chloride Channels; D002712:Chlorides / Q000494:pharmacology* / Q000502:physiology; D004159:Diphenylamine / Q000031:analogs & derivatives / Q000494:pharmacology; D004553:Electric Conductivity; D004848:Epithelium / Q000502:physiology; D005849:Gerbillinae; D007700:Kinetics; D008564:Membrane Potentials / Q000187:drug effects; D008565:Membrane Proteins / Q000502:physiology; D009579:Nitrobenzoates / Q000494:pharmacology; D012444:Saccule and Utricle / Q000187:drug effects / Q000502:physiology*
D018118:Chloride Channels; D002712:Chlorides; D008565:Membrane Proteins; D009579:Nitrobenzoates; D002034:Bumetanide; C059537:3',5-dichlorodiphenylamine-2-carboxylic acid; D012856:4-Acetamido-4'-isothiocyanatostilbene-2,2'-disulfonic Acid; C058176:5-nitro-2-(3-phenylpropylamino)benzoic acid; D004159:Diphenylamine; D017878:4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid