Skip to main content
NCBI home page
The Journal of Clinical Investigation logoLink to The Journal of Clinical Investigation
. 1991 Nov;88(5):1649–1655. doi: 10.1172/JCI115479

Evidence for direct estrogen regulation of the human gonadotropin-releasing hormone gene.

S Radovick 1, C M Ticknor 1, Y Nakayama 1, A C Notides 1, A Rahman 1, B D Weintraub 1, G B Cutler Jr 1, F E Wondisford 1
PMCID: PMC295693  PMID: 1939651

Abstract

This study is an attempt to determine whether estrogen could directly regulate human gonadotropin-releasing hormone (GnRH) gene expression. Human GnRH expression vectors were constructed by fusing various 5' flanking regions of the human GnRH gene upstream of the luciferase reporter gene (LUC) or the thymidine kinase promoter linked to the chloramphenicol acetyltransferase reporter gene (CAT). These constructs were transiently transfected into a human choriocarcinoma cell line (JEG-3) and LUC or CAT activity was measured after either no treatment or treatment with various concentrations of estradiol. A stimulatory estrogen response element (ERE) was localized to a 32-bp region between -547 and -516 bp. To determine whether estrogen receptor bound to this region of the gene, we performed DNase I footprinting using purified calf uterine estrogen receptor. DNase I footprinting demonstrates a strong footprint between -567 and -514 bp of the human GnRH gene. In addition, an avidin-biotin complex DNA-binding assay demonstrated that a biotinylated DNA fragment containing -541 to -517 bp of the human GnRH gene bound 35S-labeled estrogen receptor as well as a biotinylated DNA fragment containing the xenopus vitellogenin ERE. On the other hand, the negative control biotinylated DNA fragment derived from adenovirus 5 bound insignificant amounts of 35S-labeled estrogen receptor. Both the GnRH ERE and vitellogenin ERE bound 35S-labeled estrogen receptor with high affinity (approximately 1 nM). These data indicate that the human GnRH gene contains an ERE sufficient to mediate a stimulatory response to estrogen in heterologous cells. Based upon these data we hypothesize that the human GnRH gene might also be directly regulated by estrogen in the hypothalamus, and that this regulation may explain the GnRH hypersecretion observed at the time of the preovulatory luteinizing hormone (LH) surge.

Full text

PDF
1649

Images in this article

1653Image
on p.1653
1654Image
on p.1654

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Carmel P. W., Araki S., Ferin M. Pituitary stalk portal blood collection in rhesus monkeys: evidence for pulsatile release of gonadotropin-releasing hormone (GnRH). Endocrinology. 1976 Jul;99(1):243–248. doi: 10.1210/endo-99-1-243. [DOI] [PubMed] [Google Scholar]
  2. Cridland N. A., Wright C. V., McKenzie E. A., Knowland J. Selective photochemical treatment of oestrogen receptor in a Xenopus liver extract destroys hormone binding and transcriptional activation but not DNA binding. EMBO J. 1990 Jun;9(6):1859–1866. doi: 10.1002/j.1460-2075.1990.tb08311.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Eriksen E. F., Colvard D. S., Berg N. J., Graham M. L., Mann K. G., Spelsberg T. C., Riggs B. L. Evidence of estrogen receptors in normal human osteoblast-like cells. Science. 1988 Jul 1;241(4861):84–86. doi: 10.1126/science.3388021. [DOI] [PubMed] [Google Scholar]
  4. Glass C. K., Holloway J. M., Devary O. V., Rosenfeld M. G. The thyroid hormone receptor binds with opposite transcriptional effects to a common sequence motif in thyroid hormone and estrogen response elements. Cell. 1988 Jul 29;54(3):313–323. doi: 10.1016/0092-8674(88)90194-8. [DOI] [PubMed] [Google Scholar]
  5. Gross D. S. Effect of castration and steroid replacement on immunoreactive gonadotropin-releasing hormone in hypothalamus and preoptic area. Endocrinology. 1980 May;106(5):1442–1450. doi: 10.1210/endo-106-5-1442. [DOI] [PubMed] [Google Scholar]
  6. Klein-Hitpass L., Schorpp M., Wagner U., Ryffel G. U. An estrogen-responsive element derived from the 5' flanking region of the Xenopus vitellogenin A2 gene functions in transfected human cells. Cell. 1986 Sep 26;46(7):1053–1061. doi: 10.1016/0092-8674(86)90705-1. [DOI] [PubMed] [Google Scholar]
  7. Koller K. J., Wolff R. S., Warden M. K., Zoeller R. T. Thyroid hormones regulate levels of thyrotropin-releasing-hormone mRNA in the paraventricular nucleus. Proc Natl Acad Sci U S A. 1987 Oct;84(20):7329–7333. doi: 10.1073/pnas.84.20.7329. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Komm B. S., Terpening C. M., Benz D. J., Graeme K. A., Gallegos A., Korc M., Greene G. L., O'Malley B. W., Haussler M. R. Estrogen binding, receptor mRNA, and biologic response in osteoblast-like osteosarcoma cells. Science. 1988 Jul 1;241(4861):81–84. doi: 10.1126/science.3164526. [DOI] [PubMed] [Google Scholar]
  9. Kumar V., Chambon P. The estrogen receptor binds tightly to its responsive element as a ligand-induced homodimer. Cell. 1988 Oct 7;55(1):145–156. doi: 10.1016/0092-8674(88)90017-7. [DOI] [PubMed] [Google Scholar]
  10. Marshall J. C., Kelch R. P. Gonadotropin-releasing hormone: role of pulsatile secretion in the regulation of reproduction. N Engl J Med. 1986 Dec 4;315(23):1459–1468. doi: 10.1056/NEJM198612043152306. [DOI] [PubMed] [Google Scholar]
  11. Maurer R. A., Notides A. C. Identification of an estrogen-responsive element from the 5'-flanking region of the rat prolactin gene. Mol Cell Biol. 1987 Dec;7(12):4247–4254. doi: 10.1128/mcb.7.12.4247. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Mulvihill E. R., Palmiter R. D. Relationship of nuclear estrogen receptor levels to induction of ovalbumin and conalbumin mRNA in chick oviduct. J Biol Chem. 1977 Mar 25;252(6):2060–2068. [PubMed] [Google Scholar]
  13. Park O. K., Gugneja S., Mayo K. E. Gonadotropin-releasing hormone gene expression during the rat estrous cycle: effects of pentobarbital and ovarian steroids. Endocrinology. 1990 Jul;127(1):365–372. doi: 10.1210/endo-127-1-365. [DOI] [PubMed] [Google Scholar]
  14. Petraglia F., Vaughan J., Vale W. Steroid hormones modulate the release of immunoreactive gonadotropin-releasing hormone from cultured human placental cells. J Clin Endocrinol Metab. 1990 Apr;70(4):1173–1178. doi: 10.1210/jcem-70-4-1173. [DOI] [PubMed] [Google Scholar]
  15. Pfaff D. W. Gene expression in hypothalamic neurons: luteinizing hormone releasing hormone. J Neurosci Res. 1986;16(1):109–115. doi: 10.1002/jnr.490160111. [DOI] [PubMed] [Google Scholar]
  16. Radovick S., Wondisford F. E., Nakayama Y., Yamada M., Cutler G. B., Jr, Weintraub B. D. Isolation and characterization of the human gonadotropin-releasing hormone gene in the hypothalamus and placenta. Mol Endocrinol. 1990 Mar;4(3):476–480. doi: 10.1210/mend-4-3-476. [DOI] [PubMed] [Google Scholar]
  17. Reame N., Sauder S. E., Kelch R. P., Marshall J. C. Pulsatile gonadotropin secretion during the human menstrual cycle: evidence for altered frequency of gonadotropin-releasing hormone secretion. J Clin Endocrinol Metab. 1984 Aug;59(2):328–337. doi: 10.1210/jcem-59-2-328. [DOI] [PubMed] [Google Scholar]
  18. Roberts J. L., Dutlow C. M., Jakubowski M., Blum M., Millar R. P. Estradiol stimulates preoptic area-anterior hypothalamic proGnRH-GAP gene expression in ovariectomized rats. Brain Res Mol Brain Res. 1989 Nov;6(2-3):127–134. doi: 10.1016/0169-328x(89)90046-6. [DOI] [PubMed] [Google Scholar]
  19. Rothfeld J. M., Hejtmancik J. F., Conn P. M., Pfaff D. W. LHRH messenger RNA in neurons in the intact and castrate male rat forebrain, studied by in situ hybridization. Exp Brain Res. 1987;67(1):113–118. doi: 10.1007/BF00269459. [DOI] [PubMed] [Google Scholar]
  20. Rothfeld J., Hejtmancik J. F., Conn P. M., Pfaff D. W. In situ hybridization for LHRH mRNA following estrogen treatment. Brain Res Mol Brain Res. 1989 Nov;6(2-3):121–125. doi: 10.1016/0169-328x(89)90045-4. [DOI] [PubMed] [Google Scholar]
  21. Rudenstein R. S., Bigdeli H., McDonald M. H., Snyder P. J. Administration of gonadal steroids to the castrated male rat prevents a decrease in the release of gonadotropin-releasing hormone from the incubated hypothalamus. J Clin Invest. 1979 Feb;63(2):262–267. doi: 10.1172/JCI109298. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Seeburg P. H., Mason A. J., Stewart T. A., Nikolics K. The mammalian GnRH gene and its pivotal role in reproduction. Recent Prog Horm Res. 1987;43:69–98. doi: 10.1016/b978-0-12-571143-2.50008-3. [DOI] [PubMed] [Google Scholar]
  24. Shivers B. D., Harlan R. E., Morrell J. I., Pfaff D. W. Absence of oestradiol concentration in cell nuclei of LHRH-immunoreactive neurones. 1983 Jul 28-Aug 3Nature. 304(5924):345–347. doi: 10.1038/304345a0. [DOI] [PubMed] [Google Scholar]
  25. Shupnik M. A., Weinmann C. M., Notides A. C., Chin W. W. An upstream region of the rat luteinizing hormone beta gene binds estrogen receptor and confers estrogen responsiveness. J Biol Chem. 1989 Jan 5;264(1):80–86. [PubMed] [Google Scholar]
  26. Toranzo D., Dupont E., Simard J., Labrie C., Couet J., Labrie F., Pelletier G. Regulation of pro-gonadotropin-releasing hormone gene expression by sex steroids in the brain of male and female rats. Mol Endocrinol. 1989 Nov;3(11):1748–1756. doi: 10.1210/mend-3-11-1748. [DOI] [PubMed] [Google Scholar]
  27. Waterman M. L., Adler S., Nelson C., Greene G. L., Evans R. M., Rosenfeld M. G. A single domain of the estrogen receptor confers deoxyribonucleic acid binding and transcriptional activation of the rat prolactin gene. Mol Endocrinol. 1988 Jan;2(1):14–21. doi: 10.1210/mend-2-1-14. [DOI] [PubMed] [Google Scholar]
  28. Wondisford F. E., Farr E. A., Radovick S., Steinfelder H. J., Moates J. M., McClaskey J. H., Weintraub B. D. Thyroid hormone inhibition of human thyrotropin beta-subunit gene expression is mediated by a cis-acting element located in the first exon. J Biol Chem. 1989 Sep 5;264(25):14601–14604. [PubMed] [Google Scholar]
  29. Wray S., Grant P., Gainer H. Evidence that cells expressing luteinizing hormone-releasing hormone mRNA in the mouse are derived from progenitor cells in the olfactory placode. Proc Natl Acad Sci U S A. 1989 Oct;86(20):8132–8136. doi: 10.1073/pnas.86.20.8132. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Wray S., Zoeller R. T., Gainer H. Differential effects of estrogen on luteinizing hormone-releasing hormone gene expression in slice explant cultures prepared from specific rat forebrain regions. Mol Endocrinol. 1989 Aug;3(8):1197–1206. doi: 10.1210/mend-3-8-1197. [DOI] [PubMed] [Google Scholar]
  31. Zoeller R. T., Seeburg P. H., Young W. S., 3rd In situ hybridization histochemistry for messenger ribonucleic acid (mRNA) encoding gonadotropin-releasing hormone (GnRH): effect of estrogen on cellular levels of GnRH mRNA in female rat brain. Endocrinology. 1988 Jun;122(6):2570–2577. doi: 10.1210/endo-122-6-2570. [DOI] [PubMed] [Google Scholar]
  32. de Wet J. R., Wood K. V., DeLuca M., Helinski D. R., Subramani S. Firefly luciferase gene: structure and expression in mammalian cells. Mol Cell Biol. 1987 Feb;7(2):725–737. doi: 10.1128/mcb.7.2.725. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Clinical Investigation are provided here courtesy of American Society for Clinical Investigation

RESOURCES