Mukhopadhyay P, Geoghegan T E, Patil R V, Bhattacherjee P, Paterson C A
Department of Ophthalmology and Visual Sciences, University of Louisville School of Medicine, KY 40292, U.S.A.
Biochem Pharmacol. 1997 May 9;53(9):1249-55. doi: 10.1016/s0006-2952(97)00011-7.
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.
我们检测了三种前列腺素（PG）受体，即EP2、EP4和FP，在非色素睫状上皮细胞系（ODMCl-2）和人睫状肌（HCM）细胞中的表达情况。从ODMCl-2或HCM细胞中提取的总RNA样本，使用针对这三种PG受体各自的正义和反义引物进行逆转录聚合酶链反应（RT-PCR）。RT-PCR在ODMCl-2和HCM细胞中均产生了预测大小的对应于EP2、EP4和FP受体的DNA产物。对应于每种受体的PCR产物与特异性32P标记探针杂交，并为进一步确认，用适当的限制酶进行消化。使用EP2受体选择性激动剂布他前列素进行的药理学研究导致ODMCl-2细胞中环磷酸腺苷（cAMP）水平显著升高。PGE2和11-脱氧PGE1（分别为EP1/EP2/EP3/EP4和EP2/EP3/EP4受体激动剂）对ODMCl-2细胞中cAMP的刺激作用，被EP4受体选择性拮抗剂AH23848浓度依赖性地抑制。这些结果确凿地证明了ODMCl-2和HCM细胞中存在EP2、EP4和FP受体的mRNA和蛋白质。
