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Production in Endometrial Epithelial Cells in Vitro: Role of Cyclooxygenase-21
Département d’Ontogénie et Reproduction (E.A., P.D., M.A.F.), Centre de Recherches du Centre Hospitalier de l’Université Laval, Centre de Recherche en Biologie de la Reproduction, Ste-Foy, Québec G1V 4G2, Canada; and Département d’Obstétrique et Gynécologie (M.A.F.), Université Laval, Québec G1V 4G2, Canada
Address all correspondence and requests for reprints to: Dr. M. A. Fortier, Ontogénie et Reproduction, Centre de Recherche du Centre Hospitalier, de l’Université Laval, 2705 Boulevard Laurier, Sainte-Foy, Québec G1V 4G2, Canada. E-mail: mafortier{at}crchul.ulaval.ca
PGs are important regulators of reproductive processes. At the time of
luteolysis in vivo, PGF2
is produced by
endometrial cells, in response to oxytocin (OT). The mechanism by which
OT induces the release of PGF2 remains to be defined. We
have used 13 different cultures of bovine epithelial endometrial cells
to study the effect of OT on the regulation of PGF2 and
to identify the possible involvement of cyclooxygenases (COXs). OT
induced a dose-dependent increase of both inositol phosphates (IPs) and
[Ca2+]i concentration in epithelial cells
labeled with [3H]-myoinositol or loaded with fura-2
(using a fluorescent microscope imaging system), respectively. OT
induced a dose-dependent increase of both PGF2
production and COX-2 gene expression (as demonstrated by RT-PCR and
Northern blots). PGF2 production was increased from
13.3 ± 2.0 to 166.8 ± 22.5 ng/ml (P <
0.0001). On the other hand, COX-2/ß-actin mRNA gene expression (as
determined by densitometric analysis) was increased 5.1 ±
0.7-fold (P < 0.001) with OT (10-7
M) treatment, compared with control. Addition of
indomethacin (1 µM) and a specific COX-2 inhibitor
(NS-398, 1 µM) blocked the OT-induced PGF2
production. COX-1 and phospholipase A2 mRNA were expressed
at steady-state levels, but no effect of OT was detected on their
regulation. Combined to OT, 10 µg/ml of recombinant ovine
interferon-tau (roIFN-
) was able to decrease significantly
(P < 0.0001) the dose-dependent increase of
PGF2 production. Furthermore, partial bovine COX-1 (777
pb) and COX-2 (449 bp) cDNAs were cloned and sequenced. An homology of
83% and 97% was found in relation with rat and sheep, for COX-1,
respectively. COX-2 was found to bear 84%, 86%, and 87% of homology
in relation to rat, guinea pig, and human, respectively. Collectively,
these results demonstrate, for the first time, that COX-2 is involved
in the mechanism by which OT regulates PGF2 production
in the endometrium.
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