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a Département d'Ontogénie et Reproduction, Centre de Recherches du Centre Hospitalier de l'Université Laval (CHUL),
b Centre de Recherche en Biologie de la Reproduction (CRBR) and Département d'Obstétrique et Gynécologie, Université Laval, Ste-Foy, Québec, Canada G1V 4G2
During reproductive processes, prostaglandin (PG) E2 (PGE2) and PGF2
play important roles in which they often exert opposite effects. At the time of recognition of pregnancy in vivo, PGF2 is recognized as the luteolytic factor in ruminants and in most species including human, whereas PGE2 may exert a luteoprotective action. We have previously demonstrated that recombinant interferon-tau (rIFN-
), the embryonic signal responsible for recognition of pregnancy in ruminants, stimulated in vitro the production of PGE2 and prostaglandin-endoperoxide synthase 2 (Ptgs2; also called cyclooxygenase-2) gene expression in both epithelial and stromal endometrial cells. Since PGE2 is the major prostaglandin produced by stromal cells, the effect on Ptgs2 could explain the increase in PGE2 output. At high concentrations, however, recombinant ovine (ro) IFN- acts on epithelial cells by changing the primary PG produced from PGF2 to PGE2. This change in the primary PG produced could be explained by a decrease in PGF synthase (PGFS) activity or an increase in PGE synthase activity, or by modulation of a putative PGE2–9-ketoreductase, which converts PGE2 into PGF2. Therefore, we have investigated the regulation of the mRNAs for PGFS and PGE2-9-ketoreductase (9K-PGR), two enzymes that lead to the production of PGF2. Others have described 9K-PGR activity in uterus, ovaries, kidney, and liver of different species and have established that this enzyme could possess both 9K-PGR and 20-hydroxysteroid dehydrogenase (20-HSD) activity. Some have concluded that 9K-PGR and 20-HSD are identical enzymes. Using primers sequences chosen from homologous nucleotide sequences of published rabbit 20-HSD/9K-PGR and rat 20-HSD cDNAs, a 317-base pair (bp) fragment was amplified by reverse transcription-polymerase chain reaction (RT-PCR), cloned, and sequenced. Homologies of 83% and 78% were found with rabbit and rat 20-HSD, respectively. The presence of 20-HSD/9K-PGR and prostaglandin F synthase (PGFS) mRNA expression was studied semiquantitatively in cultured epithelial cells using RT-PCR. Stimulation of cells with roIFN-t resulted in a biphasic response, an inhibition of PGF2 production at low dose (1 ng/ml) and a stimulation of PGE2 at high dose (10 µg/ml). The increase of PGE2 was accompanied by reduced 9K-PGR and PGFS mRNA gene expression. The effect of oxytocin (OT) was also studied, and the presence of OT had no effect on either 9K-PGR or PGFS gene expression. The 20-HSD/9K-PGR transcript was also detected in other bovine tissues at different intensity (liver > kidney > testis > ovaries). We believe that the 9K-PGR and PGFS can be key enzymes in the regulation of specific PGs in the endometrium during the periimplantation period.
1 This work has been supported by Natural Sciences and Engineering Research Council of Canada (NSERC) grant #OGPIN030 (M.A.F.) and an NSERC scholarship (E.A.).
2 Correspondence: M.A. Fortier, Ontogénie et Reproduction, Centre de Recherche du Centre Hospitalier, de l'Université Laval, 2705 Boul. Laurier, Ste-Foy, PQ, Canada G1V 4G2. FAX: 418 654 2765; mafortier{at}crchul.ulaval.ca
3 Current address: Reproductive Biology Unit, Department of Obstetrics & Gynecology and Cellular & Molecular Medicine, University of Ottawa, Loeb Health Research Institute, Ottawa Civic Hospital, Ottawa, ON, Canada K1Y 4E9.
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