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Submitted on December 17, 2003
Accepted on January 7, 2004
-Hydroxysteroid Dehydrogenase Type 4 in Preovulatory Follicles and Its Potential Role in Follicular Luteinization*
Centre de recherche en reproduction animale (K.A.B., D.B., N.B., J.G.L. and J.S.) and Département de pathologie et microbiologie (M.D.), Faculté de médecine vétérinaire, Université de Montréal, Saint-Hyacinthe, Québec, Canada J2S 7C6
* To whom correspondence should be addressed.
The 17
-hydroxysteroid dehydrogenase type 4 (17-HSD4) has a unique multidomain structure, with one domain involved in estradiol-17 inactivation. The objective of the study was to investigate the regulation of 17-HSD4 during human chorionic gonadotropin (hCG)-induced ovulation/luteinization. The equine 17-HSD4 cDNA was cloned, and shown to encode a 735-amino acid protein that is highly conserved (81-87% identity) when compared with other mammalian orthologs. RT-PCR/Southern blot analyses were performed to study the regulation of 17-HSD4 transcripts in equine preovulatory follicles isolated between 0 and 39 h after hCG. Results showed the presence of basal 17-HSD4 mRNA expression before hCG treatment, but an increase was observed in follicles obtained after 24 h post-hCG (P < 0.05). Analyses of isolated preparations of granulosa and theca interna cells identified basal mRNA expression in both layers, but granulosa cells appeared as the predominant site of follicular 17-HSD4 mRNA induction. A specific polyclonal antibody was raised against a fragment of the equine protein and used to study the regulation of the 17-HSD4 protein. Immunoblots showed an increase of full-length 17-HSD4 protein in follicles after 24 h post-hCG (P < 0.05), in keeping with mRNA results. Immunohistochemical data confirmed the induction of the enzyme in follicular cells after hCG. Collectively, these results demonstrate that the gonadotropin-dependent induction of follicular luteinization is accompanied by an increase in 17-HSD4 expression. Considering the estrogen-inactivating function of 17-HSD4, its regulated expression in luteinizing preovulatory follicles appears as a potential complementary mechanism to reduce circulating levels of estradiol-17 after the LH surge.
-HSD4 •
granulosa cells •
preovulatory follicles •
luteinization •
ovary •
horse
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