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The in Vivo Control of Prostaglandin H Synthase-2 Messenger Ribonucleic Acid Expression in the Human Amnion at Parturition

Division of Obstetrics and Gynecology, John Hunter Hospital (W.B.G., W.A.W., T.Z.); and Discipline of Reproductive Medicine (R.F.J., W.B.G., W.A.W., T.Z.) and Mothers and Babies Research Center (R.F.J., C.M.M., W.B.G., W.A.W., T.Z.), University of Newcastle, Newcastle, New South Wales 2310, Australia

Address all correspondence and requests for reprints to: Dr. Tamas Zakar, Division of Obstetrics and Gynecology, Mothers and Babies Research Center, John Hunter Hospital, Locked Bag 1, Hunter Region Mail Center, Newcastle, New South Wales 2310, Australia. E-mail: . tzakar{at}mail.newcastle.edu.au

Abstract

Prostaglandin H synthase-2 (PGHS-2) activity and mRNA rise in the human amnion at late gestation, contributing to the increase in intrauterine PG production crucial for labor and delivery. In the present investigation we have determined the mechanism that controls amniotic PGHS-2 mRNA levels in vivo at term parturition.

Amnion membranes were collected after elective cesarean section (n = 20), and after spontaneous labor (n = 20). PGHS-2 relative gene transcription rates were determined by transcriptional run-on, and PGHS-2 mRNA and heterogeneous nuclear RNA (hnRNA) relative abundance were measured by quantitative real-time RT-PCR. The PGHS-2 mRNA degradation rate was determined by incubating amnion in the presence of the transcription inhibitor 5,6-dichlorobenzimidazole riboside. The dynamics of PGHS-2 hnRNA and mRNA abundance were characterized in 0- to 24-h tissue incubations.

The PGHS-2 relative gene transcription rate was a significant (P < 0.05) predictor of PGHS-2 hnRNA and mRNA abundance, and PGHS-2 hnRNA was also a predictor (P < 0.01) of PGHS-2 mRNA levels both before and after labor. Interestingly, even though PGHS-2 gene activity remained unchanged, PGHS-2 mRNA abundance increased with labor and displayed constitutive stability before and after labor. PGHS-2 mRNA levels spontaneously increased by 400% (P < 0.01) upon incubation for 24 h, whereas the transcription rate dropped by 95% during the first 2 h, then rebounded significantly between 6–24 h.

Thus, PGHS-2 mRNA abundance is transcriptionally controlled in term amnion. Labor does not increase PGHS-2 gene activity or mRNA stability. The PGHS-2 gene is probably induced before labor by a factor(s) originating in the amnion membrane, and the resulting stable mRNA accumulates progressively in the tissue throughout labor and delivery.

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