Identification of prostanoid receptors in rabbit non-pigmented ciliary epithelial cells

Preliminary ligand binding studies demonstrated that the membrane preparations of the rabbit nonpigmented ciliary epithelial cell line have 3H-prostaglandin E2 binding sites. The binding sites were specific for 3H-prostaglandin E2 as demonstrated by competition with unlabeled prostaglandin E2. The IC50 of prostaglandin E2 for the inhibition of 3H-prostaglandin E2 binding was 435 nM. The stimulation of adenylyl cyclase and phospholipase C by prostanoid receptor agonists, in rabbit non-pigmented ciliary epithelial cells resulted in the formation of either cyclic AMP or inositol phosphates. Prostaglandin E2 and 16-16-dimethyl prostaglandin E2 (both are EP1, EP2, EP3 and EP4 receptor agonists). 11-deoxy prostaglandin E1 (EP2, EP3 and EP4 receptor agonist), butaprost (EP2 receptor agonist), and prostaglandin D2 (DP receptor agonist) stimulated the formation of cyclic AMP in a dose-dependent manner. Maximal stimulation occurred between 1.25 and 2.5 microM for prostaglandin E2 and 16,16-dimethyl prostaglandin E2 and between 10 and 20 microM for 11-deoxy prostaglandin E1 and prostaglandin D2. Prostaglandin E2 and 16,16-dimethyl prostaglandin E2 were more potent (EC50 of 0.25 microM and 0.42 microM respectively) than 11-deoxy prostaglandin E1, butaprost or prostaglandin D2. The formation of cyclic AMP by prostaglandin D2 was inhibited by BW868C, a highly selective DP receptor antagonist. 17-phenyl trinor prostaglandin E2, prostaglandin F2 alpha and U46619, the EP1, FP and TP receptor agonists, respectively stimulated phospholipase C (as measured by the formation of total inositol phosphates) in a dose-dependent manner. The agonists 11-deoxy prostaglandin E1 and butaprost coupled to adenylyl cyclase via guanine nucleotide binding protein, G8, did not increase the turnover of inositol phosphates. The results of the present study suggest that rabbit non-pigmented ciliary epithelial cells express EP1, EP2, DP, FP and TP receptors.