Down-regulation of messenger ribonucleic acid encoding an importer of sulfoconjugated steroids during human chorionic gonadotropin-induced follicular luteinization in vivo

• Published in: The Journal of steroid biochemistry and molecular biology Vol. 103; no. 1; pp. 10 - 19
• Main Authors: Brown, Kristy A., Bouchard, Nadine, Lussier, Jacques G., Sirois, Jean
• Format: Journal Article
• Language: English
• Published: Oxford Elsevier Ltd 2007; Elsevier Science
• Subjects: Amino Acid Sequence; Animals; Base Sequence; Biological and medical sciences; Chorionic Gonadotropin - pharmacology; Corpus Luteum - metabolism; Down-Regulation; Female; Fundamental and applied biological sciences. Psychology; Granulosa cells; Hormone metabolism and regulation; Horse; Horses; Luteinization; Mammalian female genital system; Molecular Sequence Data; Ovarian Follicle - metabolism; Ovary; Preovulatory follicles; RNA, Messenger - metabolism; Sequence Homology, Amino Acid; SLCO2B1; Steroid sulfatase; Steryl-Sulfatase - metabolism; Tissue Distribution; Transcription, Genetic; Vertebrates: reproduction; Preovulatory follicles; Ovary; Steroid sulfatase; SLCO2B1; Horse; Granulosa cells; Luteinization; Human; Steroid; Import; Gonadotropin; Gene expression; Placental hormone; Human chorionic gonadotrophin; Vertebrata; Mammalia; Animal; Female genital system; Perissodactyla; Granulosa; Molecular cloning; Ovarian follicle; Sulfoconjugation; Ungulata
• DOI: 10.1016/j.jsbmb.2006.07.005
• ISSN: 0960-0760

Members of the organic anion transporting polypeptide (SLCO/OATP) superfamily are capable of importing anionic compounds across the lipid bilayer in a sodium-independent manner. Member 2B1 has been shown to transport few substrates, two of which are dihydroepiandrosterone-3-sulfate (DHEA-S) and estrone-3-sulfate. Steroid sulfatase (STS) catalyses the hydrolysis of these steroids into their unconjugated counterparts. The objective of this study was to investigate the regulation of SLCO2B1 and STS mRNAs during human chorionic gonadotropin (hCG)-induced ovulation/luteinization. The equine SLCO2B1 cDNA was cloned and shown to encode a 709-amino acid protein (OATP2B1) that is highly conserved when compared to mammalian orthologs. RT-PCR/Southern blot analyses were performed to study the regulation of SLCO2B1 and STS transcripts in equine preovulatory follicles isolated between 0 and 39 h after hCG treatment. Results showed high levels of SLCO2B1 mRNA expression before hCG, with a marked decrease observed in follicles obtained 24–39 h post-hCG ( P < 0.05). Analyses of isolated granulosa and theca interna cells identified high mRNA expression in both cell types prior to hCG treatment, with granulosa cells showing a more rapid SLCO2B1 mRNA down-regulation. No significant change in STS mRNA was observed in intact follicle walls. However, when both cell types were isolated, a significant decrease in STS mRNA was observed in granulosa cells 24–39 h post-hCG. Collectively, these results demonstrate that the hCG-dependent induction of follicular luteinization is accompanied by the down-regulation of SLCO2B1 and STS transcripts. Considering that OATP2B1 can import sulfoconjugated DHEA and estrogens, and that STS can remove the sulfonate moiety from these steroids, their down-regulation in luteinizing preovulatory follicles may provide an additional biochemical basis for the decrease in ovarian 17β-estradiol biosynthesis after the LH surge.