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, EP,, EP,, and FP, in a nonpigmented ciliary epithelial cell line (ODM Cl-2) and in human ciliary muscle (HCM) cells. Total RNA preparations from either ODMCl-2 or HCM cells were subjected to reverse transcripti on-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 recept or were hybridized with specific P-32-labeled probes and, for further confirmation, digested with appropriate restriction enzymes. Pharmacol ogical studies with the EP, receptor-selective agonist butaprost resul ted in a significant increase in the cyclic AMP lever in ODMCl-2 cells . The stimulation of cyclic AMP in ODMCl-2 cells by PGE(2) and 11-deox y PGE(1), the respective EP1/EP2/EP3/EP4 and EP2/EP3/EP4 receptor agon ists, was concentration dependently inhibited by the EP4 receptor-sele ctive antagonist AH23848. These results conclusively demonstrate the p resence of both mRNA and protein for EP2,EP4, and FP receptors in ODMC l-2 and HCM cells. (C) 1997 Elsevier Science Inc.