Effect of Heptanol on the Short Circuit Currents of Cornea and Ciliary Body Demonstrates Rate Limiting Role of Heterocellular Gap Junctions in Active Ciliary Body Transport

Abstract

Rabbit ciliary body and cornea were mounted in Ussing-type chambers in Tyrode's under voltage clamp and the effects of heptanol, a gap junction inhibitor, on the short circuit current generated by each of the respective epithelia were determined. Studies were carried out either in control conditions or following amphotericin B permeabilization of either the basolateral membrane of the nonpigmented epithelium of the ciliary body or the apical membrane of the corneal epithelium, respectively. Previous studies have shown that, following these permeabilizations, short circuit currents are established, reflecting aqueous (or tear) -to-serosa Na+fluxes, and that Na+translocation through gap junctions connecting the individual layers of these tissues constitutes the major rate limiting step. Heptanol inhibited most of the short circuit current of the amphotericin B-modified ciliary body and cornea and of the unmodified ciliary body epithelium (control). In all these cases, the apparentIC50was about 0.8M. In the unmodified corneal epithelium, where ion translocation across the apical membrane constitutes the main rate limiting step for active secretion, 0.4 or 0.8 mMheptanol induced short circuit current increases; partial inhibition was observed only at high concentrations known to cause maximal inhibition of junctional permeability. Heptanol also enhanced the volume regulatory decrease of cultured human NPE cells, a process dependent on cell swelling-induced stimulation of Cl−and K+permeabilities. Combined with our previous results demonstrating the lack of heptanol effects on other epithelial functions, these data suggest that the effect of heptanol on the active ciliary body transepithelial transport is primarily due to inhibition of the nonpigmented-pigmented junctional path and that this path is a potential site of rate limitation for the secretory process.