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J. Biol. Chem., Vol. 276, Issue 36, 34323-34330, September 7, 2001
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From the Centre de recherche en reproduction animale and the
Département de biomédecine vétérinaire,
Faculté de médecine vétérinaire,
Université de Montréal, C.P. 5000, Saint-Hyacinthe, Québec J2S 7C6, Canada
Prostaglandin E2
(PGE2) is thought to be an ultimate prostaglandin effector
during the ovulatory process, and the objectives of this study were to
clone bovine PGE synthase (PGES) and to characterize its regulation by
gonadotropins in preovulatory follicles in vivo. The bovine
PGES complementary DNA (cDNA) was shown to contain a
5'-untranslated region of eight base pairs (bp), an open reading frame
of 462 bp and a 3'-untranslated region of 406 bp. The putative bovine
PGES open reading frame encodes a 153-amino acid protein that is 85, 78, and 78% identical to the human, rat, and mouse PGES homologs,
respectively. The regulation of PGES during ovulation was
studied using three different models in vivo: 1) human
chorionic gonadotropin (hCG)-induced ovulation during a normal estrous
cycle; 2) hCG-induced ovulation following ovarian hyperstimulation; and
3) spontaneous ovulation during natural estrus. Results from
semi-quantitative reverse transcription-polymerase chain
reaction/Southern blotting analyses showed that the hCG/luteinizing hormone surge caused a significant increase in PGES mRNA. Levels of
PGES transcripts were low or undetectable prior to hCG/luteinizing hormone but increased markedly 18-24 h after hCG in models 1 and 2, and 18-24 h after the onset of natural estrus in model 3 (p < 0.05). Analyses on isolated preparations of
granulosa and theca interna cells indicated that the granulosa cell
layer was the predominant site of follicular PGES expression. The
regulation of the protein was studied in the same models using a
specific antibody raised against a fragment of bovine protein ( The nucleotide sequence(s) reported in this paper has been submitted to the DDBJ/GenBankTM/EBI Data Bank with accession number(s) AY032727.
Molecular Cloning and Induction of Bovine Prostaglandin E
Synthase by Gonadotropins in Ovarian Follicles Prior to Ovulation
in Vivo*
§
PGES;
from Glu49 to Val146). Results from
immunoblots showed an induction of bovine PGES (Mr = 17,000) 18-24 h after hCG treatment or
onset of estrus (p < 0.05). The protein was detected
in extracts of granulosa cells but not in theca interna. Collectively,
these results demonstrate that the ovulatory process is associated with
a gonadotropin-dependent induction of PGES in granulosa
cells of ovarian follicles in vivo, thus establishing for
the first time the regulation of the enzyme in a physiological context.
*
This work was supported by Canadian Institutes of Health
Research (CIHR) Grant MT-13190 (to J. S.) and Fonds pour la Formation de Chercheurs et l'Aide à la Recherche Grant 99-ER-3016 (to
J. G. L. and J. S.).The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
Supported by a CIHR Investigator Award.
§
To whom correspondence should be addressed: Tel.: 450-773-8521 (ext. 8542); Fax: 450-778-8103; E-mail:
siroisje@medvet.umontreal.ca.
Copyright © 2001 by The American Society for Biochemistry and Molecular Biology, Inc.
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