Transepithelial transport of ascorbic acid by the isolated intact ciliary epithelial bilayer of the rabbit eye

The purpose of these experiments was to test whether the isolated intact epithelial bilayer derived from rabbit ciliary processes transports ascorbic acid and to what degree the transport rate in vitro corresponds to the in vivo process. The intact ciliary epithelial bilayer of the rabbit eye isolated by perfusion was mounted in a particularly constructed Ussing type chamber. Fluxes were measured by additions of 14C ascorbate to the hemichamber on either the pigmented epithelium (PE) or the nonpigmented epithelium (NPE) side where equal concentrations of ascorbate from .02 to 2.0 mM were present. Samples were taken at intervals thereafter and counted in a liquid scintillation counter. The experiments were done under short circuit conditions to avoid the possible influence of fluctuating currents upon the movement of ascorbate. Like the earlier iris ciliary body preparations, separation of ascorbate fluxes is also done by the isolated intact ciliary epithelial bilayer, and the transport of ascorbic acid proceeds by saturation kinetics. The uptake process is accomplished entirely by the pigmented epithelium (PE). The Km of the process is 0.97 mM, and Vmax was valued at 130 nM/L/hr. Thus, assuming an aqueous flow rate of 2 microliters/min, the transfer of ascorbic acid across the bilayer occurs at a rate required to maintain the ordinary millimolar concentration of ascorbic acid in the aqueous humor found in vivo.