Detection of EP2, EP4, and FP receptors in human ciliary epithelial and ciliary muscle cells.
We have examined the expression of three prostaglandin (PG) receptors, EP2, EP4, and FP, in a nonpigmented ciliary epithelial cell line (ODMCl-2) and in human ciliary muscle (HCM) cells. Total RNA preparations from either ODMCl-2 or HCM cells were subjected to reverse transcription-polymerase chain reaction (RT-PCR) with sense and antisense primers for each of the three PG receptors. The RT-PCR generated DNA products of predicted sizes corresponding to the EP2, EP4, and FP receptors in both ODMCl-2 and HCM cells. PCR products corresponding to each receptor were hybridized with specific 32P-labeled probes and, for further confirmation, digested with appropriate restriction enzymes. Pharmacological studies with the EP2 receptor-selective agonist butaprost resulted in a significant increase in the cyclic AMP level in ODMCl-2 cells. The stimulation of cyclic AMP in ODMCl-2 cells by PGE2 and 11-deoxy PGE1, the respective EP1/EP2/EP3/EP4 and EP2/EP3/EP4 receptor agonists, was concentration-dependently inhibited by the EP4 receptor-selective antagonist AH23848. These results conclusively demonstrate the presence of both mRNA and protein for EP2, EP4, and FP receptors in ODMCl-2 and HCM cells.
Mukhopadhyay
P
P
Department of Ophthalmology and Visual Sciences, University of Louisville School of Medicine, KY 40292, U.S.A.
Geoghegan
T E
TE
Patil
R V
RV
Bhattacherjee
P
P
Paterson
C A
CA
D000527:Alprostadil / Q000031:analogs & derivatives / Q000494:pharmacology; D002478:Cells, Cultured / Q000378:metabolism; D002924:Ciliary Body / Q000378:metabolism*; D015232:Dinoprostone / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D004848:Epithelium / Q000378:metabolism; D006801:Humans; D010857:Pigment Epithelium of Eye / Q000378:metabolism; D011982:Receptors, Prostaglandin / Q000032:analysis* / Q000145:classification; D018078:Receptors, Prostaglandin E / Q000032:analysis* / Q000145:classification; D058309:Receptors, Prostaglandin E, EP4 Subtype
C545650:PTGER4 protein, human; D011982:Receptors, Prostaglandin; D018078:Receptors, Prostaglandin E; D058309:Receptors, Prostaglandin E, EP4 Subtype; C034671:prostaglandin F2alpha receptor; D000527:Alprostadil; D015232:Dinoprostone; C034008:11-deoxyprostaglandin E1