Expression of genes involved in prostaglandin E2 and progesterone production in bovine cumulus-oocyte complexes during in vitro maturation and fertilization.
Prostaglandin E(2) (PGE(2)) and progesterone appear to be critical mediators of cumulus expansion and the resumption of oocyte meiosis. The aim of this study was to identify the types of prostaglandin E synthase (PTGES) expressed in the bovine cumulus-oocyte complex (COC), to characterize their temporal expression during the periconceptional interval using an in vitro model of maturation (IVM) and fertilization (IVF), and to compare their expression with the level of steroidogenic gene expression. Real-time RT-PCR analysis revealed that enzymes related to the PGE(2) biosynthesis pathway were mainly expressed during IVM. Transcripts encoding PTGES1-3 were detected in bovine COCs. Only the expression of PTGES1 significantly increased during IVM whereas that of PTGES2 and PTGES3 remained unchanged. The induction of PTGES1 expression paralleled the induction of prostaglandin G/H synthase-2 (PTGS2) expression and the amounts of PGE(2) secreted by maturing COCs. Concomitantly, cholesterol side chain cleavage cytochrome P450 expression was significantly upregulated in maturing COCs and the high level of expression persisted in fertilized COCs. The expression of the StAR protein remained constant during IVM and then decreased significantly during IVF. Expression of the progesterone catabolic-related enzyme, 20alpha-hydroxysteroid dehydrogenase significantly decreased throughout the periconceptional interval. This was associated with a rising level of progesterone released by COCs in the culture media. In conclusion, our results suggest that the periconceptional differentiation of the bovine COC includes the transient induction of PGE(2) biosynthetic activity via the PTGS2/PTGES1 pathway during the maturation period and the increasing ability to produce progesterone from the immature to the fertilized stages.
Nuttinck
Fabienne
F
ENVA, UMR 1198, Biologie du Développement et Reproduction, 7 av du Général De Gaulle, Maisons-Alfort F-94704, France. fabienne.nuttinck@jouy.inra.fr
Marquant-Le Guienne
Brigitte
B
Clément
Laetitia
L
Reinaud
Pierrette
P
Charpigny
Gilles
G
Grimard
Bénédicte
B
D015089:20-Hydroxysteroid Dehydrogenases / Q000235:genetics; D000818:Animals; D002417:Cattle; D002478:Cells, Cultured; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics; D015232:Dinoprostone / Q000096:biosynthesis*; D005260:Female; D005307:Fertilization in Vitro; D015870:Gene Expression; D018507:Gene Expression Regulation, Developmental; D019746:Intramolecular Oxidoreductases / Q000235:genetics; D009865:Oocytes / Q000502:physiology*; D009866:Oogenesis / Q000235:genetics*; D011374:Progesterone / Q000096:biosynthesis*; D000071876:Prostaglandin-E Synthases; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods
D011374:Progesterone; D003577:Cytochrome P-450 Enzyme System; D015089:20-Hydroxysteroid Dehydrogenases; D019746:Intramolecular Oxidoreductases; D000071876:Prostaglandin-E Synthases; D015232:Dinoprostone