Pharmacological and molecular biological (RT-PCR) characterization of functional TP prostanoid receptors in immortalized human non-pigmented ciliary epithelial cells.
Immortalized human non-pigmented ciliary epithelial (NPE) cells (ODM-2) were shown to express the mRNA for the prostanoid TPalpha but not the TPbeta receptor using reverse transcription-polymerase chain reaction (RT-PCR). These TPalpha receptors were coupled to phospholipase C (PLC) and, thus, promoted phosphoinositide (PI) turnover. TP receptor agonists yielded the following potencies (EC50S) in the PI turnover assays: I-BOP = 8.2 +/- 1.1 nM; carbocyclic TA2 = 87.5 +/- 25.3 nM; U-44069 = 1.16 +/- 0.32 microM; U-46619 = 1.2 +/- 0.2 microM (n = 4-17). Agonists selective for other prostanoid receptor subtypes (e.g., fluprostenol and sulprostone) were inactive. The agonist effects of U-44619 and I-BOP were potently blocked, in an apparent non-competitive manner (ki = 53.9 +/- 12 nM; pA2s = 7.6-7.8; pKbs = 7.38), by the TP receptor-selective antagonist, SQ29,548, but were unaffected by other prostanoid receptor antagonists (e.g., AH6809, AL-8810). The PLC inhibitor (U73122) inhibited U-46619-induced PI turnover (IC50 = 4.3 +/- 0.6 microM). The functional potencies of the compounds stimulating or inhibiting the TP receptor-mediated PI turnover in the NPE cells correlated well with the TP receptor binding affinities of these compounds at human platelet TP receptors (r = 0.98). These studies have shown the presence of the mRNA for and the expression of functional TPalpha receptors coupled to PLC in human NPE cells. The TPalpha receptors on NPE cells may be responsible for inhibiting aqueous humor production and may help explain the intraocular pressure-lowering effects of certain TP agonists.
Sharif
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Molecular Pharmacology Unit, Alcon Research Ltd., Fort Worth, Texas 76134-2099, USA. naj.sharif@alconlabs.com
Senchyna
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Xu
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D017398:Alternative Splicing; D002461:Cell Line, Transformed; D002924:Ciliary Body / Q000166:cytology / Q000378:metabolism*; D004847:Epithelial Cells / Q000378:metabolism; D006801:Humans; D010716:Phosphatidylinositols / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D018078:Receptors, Prostaglandin E / Q000235:genetics; D058307:Receptors, Prostaglandin E, EP2 Subtype; D017482:Receptors, Thromboxane / Q000235:genetics / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction
C545644:PTGER2 protein, human; D010716:Phosphatidylinositols; D012333:RNA, Messenger; D018078:Receptors, Prostaglandin E; D058307:Receptors, Prostaglandin E, EP2 Subtype; D017482:Receptors, Thromboxane