[Induction and properties of nitric oxide synthase in rat retinal pigment epithelial cells in culture].
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.


CONCLUSIONS
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.
Furukawa
M
M
Department of Ophthalmology, Aichi Medical University, Japan.
Iwaki
M
M
Ogino
N
N
Liu
J
J
Nishida
S
S
Mandai
M
M
D000594:Amino Acid Oxidoreductases / Q000096:biosynthesis*; D000818:Animals; D001483:Base Sequence; D002478:Cells, Cultured; D004790:Enzyme Induction; D008969:Molecular Sequence Data; D009569:Nitric Oxide / Q000096:biosynthesis; D019001:Nitric Oxide Synthase; D010857:Pigment Epithelium of Eye / Q000201:enzymology*; D051381:Rats
D009569:Nitric Oxide; D019001:Nitric Oxide Synthase; D000594:Amino Acid Oxidoreductases