Human chorionic gonadotropin-dependent up-regulation of genes responsible for estrogen sulfoconjugation and export in granulosa cells of luteinizing preovulatory follicles.
Estrogen sulfotransferase (EST) is responsible for the sulfoconjugation of estrogens, thereby changing their physical properties and preventing their action via the estrogen receptors. These sulfoconjugated steroids no longer diffuse freely across the lipid bilayer; instead, they are exported by members of the ATP-binding cassette family, such as ABCC1. The objective of this study was to investigate the regulation of EST and ABCC1 during human chorionic gonadotropin (hCG)-induced ovulation/luteinization. The transcripts for EST and ABCC1 were cloned by RT-PCR, and the regulation of their mRNAs was studied in preovulatory follicles obtained during estrus at 0, 12, 24, 30, 33, 36, and 39 h after hCG. Results obtained from RT-PCR/Southern blot analyses showed significant changes in steady-state levels of both EST and ABCC1 mRNA after hCG treatment (P < 0.05). In granulosa cells, a significant increase in EST transcript was observed 30-39 h after hCG. Similarly, ABCC1 transcript levels were induced in granulosa cells 12-39 h after hCG. In contrast, no significant changes in either EST or ABCC1 were detected in theca interna samples after hCG. The increase in EST and ABCC1 transcripts observed in granulosa cells was reflected in preparations of intact follicle walls, suggesting that the granulosa cell layer contributes the majority of EST and ABCC1 expression in preovulatory follicles. The present study demonstrates that follicular luteinization is accompanied not only by a decrease in 17 beta-estradiol biosynthesis but also by an increase in expression of genes responsible for estrogen inactivation and elimination from granulosa cells, such as EST and ABCC1, respectively.
Brown
Kristy A
KA
Centre de Recherche en Reproduction Animale and Département de Biomédecine Vétérinaire, Faculté de Médecine Vétérinaire, Université de Montréal, Saint-Hyacinthe, Québec, Canada J2S 7C6.
Doré
Monique
M
Lussier
Jacques G
JG
Sirois
Jean
J
D000595:Amino Acid Sequence; D000818:Animals; D015139:Blotting, Southern; D002417:Cattle; D006063:Chorionic Gonadotropin / Q000494:pharmacology*; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000737:chemistry; D004967:Estrogens / Q000378:metabolism*; D005260:Female; D005498:Follicular Phase; D006107:Granulosa Cells / Q000737:chemistry / Q000378:metabolism*; D006736:Horses; D006801:Humans; D034461:Luteinization; D008969:Molecular Sequence Data; D027425:ATP-Binding Cassette, Sub-Family C Proteins / Q000235:genetics; D006080:Ovarian Follicle / Q000737:chemistry / Q000166:cytology; D012333:RNA, Messenger / Q000032:analysis / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016415:Sequence Alignment; D013431:Sulfates / Q000378:metabolism; D015238:Sulfotransferases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D015854:Up-Regulation / Q000187:drug effects*
D006063:Chorionic Gonadotropin; D018076:DNA, Complementary; D004967:Estrogens; D027425:ATP-Binding Cassette, Sub-Family C Proteins; D012333:RNA, Messenger; D013431:Sulfates; D015238:Sulfotransferases; C020122:estrone sulfotransferase; C421109:multidrug resistance-associated protein 1