PMID: 11923260Mar 30, 2002Paper

Net glutathione secretion across primary cultured rabbit conjunctival epithelial cell layers

Investigative Ophthalmology & Visual Science
Hovhannes J GukasyanRam Kannan

Abstract

Metabolism and transport of glutathione (GSH), the endogenous thiol antioxidant, in conjunctival tissue to date are poorly understood. The purpose of the present study was to define transport characteristics of GSH in primary cultured rabbit conjunctival epithelial cells (RCECs). RCECs were grown on membrane filters to exhibit tight barrier properties (transepithelial electrical resistance, TEER, approximately 1 k(Omega)/cm(2)). Uptake, efflux, and transepithelial transport of GSH were determined in the presence or absence of extracellular Na(+) under conditions of inhibition of GSH biosynthesis and degradation. Uptake was determined at 15 minutes after instillation of (3)H-GSH to the apical or basolateral bathing fluid. GSH efflux was estimated from the time course of release of prebiosynthesized (35)S-GSH. Transepithelial transport was assessed by instillation of (3)H-GSH in either the apical or basolateral bathing fluid, followed by sampling from respective contralateral sides. Apical uptake and efflux showed Na(+) dependency up to 65%. GSH uptake in the initial 15 minutes was linear in the presence of 1 mM GSH (labeled and unlabeled) in Na(+)-containing buffer. The uptake rate was higher from the apical fluid than from the ...Continue Reading

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