Unlabelled: We investigated the production of nitric oxide (NO) and nitric oxide synthase (NOS) by rat retinal pigment epithelial cells (RPE) in culture.
Methods: RPE was isolated from the eyes of 7-10 day-old rats and cultured in Dulbecco's modified Eagle medium with 10% fetal calf serum. NO2- concentration in the culture medium was determined with the Griess reaction. NOS was assayed by measuring 3H citrulline as the reaction product. RT-PCR was performed on the isolated RNA fractions from RPE.
Results: Rat RPE produced NOS with the combined stimulation of the bacterial endotoxin and cytokines including interferon gamma. PCR experiments showed that stimulated RPE expressed a gene product corresponding to inducible-type NOS. The enzyme required arginine, NADPH, and tetrahydrobiopterin for the reaction. The specific activity of the crude RPE extract was approximately 270 pmol/min/mg.
Conclusion: Like bovine RPE, rat RPE produced NOS by the immunological stimulation. Induced NOS and the product NO may play important roles in inflammation of the eye.
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