The disposition and bioavailability of 35S-GSH from 35S-GSSG in BSS PLUS in rabbit ocular tissues

The purpose of this study was to evaluate the biodistribution and uptake of 35S-GSH into intraocular tissues following the administration of BSS PLUS containing 35S-GSSG by either an anterior chamber or intravitreal injection. This study evaluated the disposition and uptake of the 35S-radiolabel, the intracellular concentrations of 35S-GSH from extracellular 35S-GSSG, and the percentage of 35S-GSH to the total cellular GSH pool. Glutathione was analyzed by high-performance liquid chromatography (HPLC) using fluorescence detection after derivitizing the thiols in situ with monobromobimane. The effluent from the GSH peak was then collected for measurement of 35S-GSH. After an anterior chamber injection of 35S-BSS PLUS, 35S-radioactivity rapidly disappeared from the aqueous humor between 0.5 and 2 hours; corneal 35S-radioactivity remained constant over time. 35S-GSH was detected in the iris and ciliary body. However, in the cornea, 35S-GSH became the predominant radioactive thiol in the stroma, endothelium, and epithelium; the corneal stroma appeared to be a possible GSH reservoir for the adjacent corneal layers. After an intravitreal injection, 35S-radioactivity slowly decreased in the vitreous humor but was readily taken up by the tissues of the posterior segment, especially the retina and choroid, which showed the greatest concentrations of 35S-GSH of all tissues studied. The data from this study demonstrate that 35S-GSSG in BSS PLUS is metabolized and taken up by ocular cells and that 35S-GSH becomes incorporated into the intracellular GSH pool of ocular tissues.