Cl− secretory effects of EBIO in the rabbit conjunctival epithelium

Abstract

Experiments were conducted to determine whether the Cl− secretagogue, 1-ethyl-2-benzimidazolinone (EBIO), stimulates Cl− transport in the rabbit conjunctival epithelium. For this study, epithelia were isolated in an Ussing-type chamber under short-circuit conditions. The effects of EBIO on the short-circuit current ( Isc) and transepithelial resistance ( Rt) were measured under physiological conditions, as well as in experiments with altered electrolyte concentrations. Addition of 0.5 mM EBIO to the apical bath stimulated the control Isc by 64% and reduced Rt by 21% ( P < 0.05; paired data). Under Cl−-free conditions, Isc stimulation using EBIO was markedly attenuated. In the presence of an apical-to-basolateral K+ gradient and permeabilization of the apical membrane, the majority of the Isc reflected the transcellular movement of K+ via basolateral K+ channels. Under these conditions, EBIO in combination with A23187 elicited nearly instantaneous 60–90% increases in Isc that were sensitive to the calmodulin antagonist calmidazolium and the K+ channel blocker tetraethyl ammonium. In the presence of an apical-to-basolateral Cl− gradient and nystatin permeabilization of the basolateral aspect, EBIO increased the Cl−-dependent Isc, an effect prevented by the channel blocker glibenclamide (0.3 mM). The latter compound also was used to determine the proportion of EBIO-evoked unidirectional 36Cl− fluxes in the presence of the Cl− gradient that traversed the epithelium transcellularly. Overall, EBIO activated apical Cl− channels and basolateral K+ channels (presumably those that are Ca2+ dependent), thereby suggesting that this compound, or related derivatives, may be suitable as topical agents to stimulate fluid transport across the tissue in individuals with lacrimal gland deficiencies.