The expression of aromatase in gonadotropes is regulated by estradiol and gonadotropin-releasing hormone in a manner that differs from the regulation of luteinizing hormone.
The role of estrogens is dual: they suppress basal expression of gonadotropins and enhance GnRH responsiveness at the time of the LH surge. Estrogens are synthesized by cytochrome P450 aromatase (P450arom), encoded by the cyp19 gene. We focused on the cyp19 gene in rat and showed that it is expressed in gonadotropes through promoters PII and PI.f, using RT-PCR and dual fluorescence labeling with anti-P450arom and -LH antibodies. Real-time PCR quantification revealed that aromatase mRNA levels varied during the estrous cycle and were significantly increased after ovariectomy. This effect is prevented by estradiol (E2) as well as GnRH antagonist administration, suggesting that GnRH may mediate the steroid effect. Interestingly, the long-acting GnRH agonist that induces LH desensitization does not modify aromatase expression in ovariectomized rats. Administration of E2 in ovariectomized rats receiving either GnRH agonist or GnRH antagonist clearly demonstrated that E2 also reduces cyp19 expression at the pituitary level. The selective estrogen receptor-alpha ligand propyl pyrazole triol and the selective estrogen receptor-beta ligand diarylpropionitrile both mimic the E2 effects. By contrast, propyl pyrazole triol reduces LH beta expression whereas diarylpropionitrile does not. In addition, using transient transfection assays in an L beta T2 gonadotrope cell line, we provided evidence that GnRH agonist stimulated, in a dose-dependant manner, cyp19 promoters PII and PI.f and that E2 decreased the GnRH stimulation. In conclusion, our data demonstrate that GnRH is an important signal in the regulation of cyp19 in gonadotrope cells. Both common and specific intracellular factors were responsible for dissociated variations of LH beta and cyp19 expression.
Galmiche
Guillaume
G
Département Génétique et Reproduction, UFR de médecine, F-14033 Caen, France. kottler-ml@chu-caen.fr
Richard
Nicolas
N
Corvaisier
Sophie
S
Kottler
Marie-Laure
ML
D000818:Animals; D001141:Aromatase / Q000235:genetics*; D015153:Blotting, Western; D004958:Estradiol / Q000494:pharmacology*; D047628:Estrogen Receptor alpha / Q000502:physiology; D047629:Estrogen Receptor beta / Q000502:physiology; D030762:Estrous Cycle; D005260:Female; D005455:Fluorescent Antibody Technique; D005786:Gene Expression Regulation / Q000187:drug effects*; D007987:Gonadotropin-Releasing Hormone / Q000037:antagonists & inhibitors / Q000494:pharmacology*; D037101:Luteinizing Hormone, beta Subunit / Q000235:genetics*; D008297:Male; D010052:Ovariectomy; D010902:Pituitary Gland / Q000201:enzymology*; D011401:Promoter Regions, Genetic / Q000235:genetics; D012333:RNA, Messenger / Q000032:analysis; D051381:Rats; D017207:Rats, Sprague-Dawley; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014162:Transfection
D047628:Estrogen Receptor alpha; D047629:Estrogen Receptor beta; D037101:Luteinizing Hormone, beta Subunit; D012333:RNA, Messenger; D007987:Gonadotropin-Releasing Hormone; D004958:Estradiol; D001141:Aromatase