Active transport of ascorbate across the isolated rabbit ciliary epithelium

Abstract

The transepithelial transport of ascorbate across the isolated rabbit ciliary epithelium (CE) was investigated. Unidirectional 14C-ascorbate fluxes were measured in the presence of equal concentrations of ascorbate on both sides of the tissue within the range of 0.025 to 1 mM. The blood to aqueous (Bl----Aq) flux increased from 6 to 95 nmoles/hr and showed nonlinearity and saturation. The aqueous to blood (Aq----Bl) flux increased, for the same range, from 0.5 to 23 nmoles/hr in a linear fashion. The permeability calculated from the Aq----Bl flux was similar to the CE permeability for mannitol suggesting that the Aq----Bl flux is mainly paracellular. The flux ratio Bl----Aq/Aq----Bl was between 4 to 12. Anoxia, ouabain and low Na+ in the media inhibited the Bl----Aq flux indicating that the transport system requires energy and a Na+ gradient. 3-O-methyl-D-glucose, D-isoascorbic acid and phlorizin also inhibited the Bl----Aq flux, suggesting that ascorbate and glucose may share a common carrier mechanism. Although the isolated CE preparation was clearly capable of flux separation and active transport, the rate of ascorbate transport measured in vitro is insufficient to maintain the aqueous ascorbate concentration observed in vivo.