Skip to main content
PMC home page
Biochemical Journal logoLink to Biochemical Journal
. 1995 Jul 1;309(Pt 1):325–329. doi: 10.1042/bj3090325

Mechanism of action of gonadotropin-releasing hormone upon gonadotropin alpha-subunit mRNA levels in the alpha T3-1 cell line: role of Ca2+ and protein kinase C.

D Ben-Menahem 1, Z Shraga-Levine 1, P L Mellon 1, Z Naor 1
PMCID: PMC1135837  PMID: 7542447

Abstract

Addition of [D-Trp6]gonadotropin-releasing hormone (GnRHa) to alpha T3-1 cells induced a very rapid response upon gonadotropin alpha-subunit mRNA which was detected after 30-60 min and was abolished by pretreatment with actinomycin D. A similar response was obtained with the protein kinase C (PKC) activator phorbol 12-myristate 13-acetate (PMA), or the Ca2+ ionophore, ionomycin. GnRHa (10 nM) also stimulated a secondary rise in alpha-subunit mRNA levels between 12 and 24 h of incubation. No additivity was obtained (at 60 min) upon the combined addition of GnRHa and PMA, GnRHa and ionomycin, or PMA and ionomycin. The effect of GnRHa upon alpha-subunit mRNA was blocked by the PKC inhibitors staurosporine or GF 109203X. Down-regulation of endogenous PKC activity resulted in inhibition of the stimulatory effect of gonadotropin-releasing hormone (GnRH), PMA and ionomycin. Removal of extra-cellular Ca2+ abolished the effect of GnRHa and PMA upon alpha-subunit mRNA levels. Interestingly PMA and ionomycin had no effect on alpha-subunit mRNA levels at 24 h of incubation; however, the combined addition of the drugs mimicked the late phase of GnRHa (10 nM) action. The data provide evidence that PKC and Ca2+ are involved in mediating the early and the late responses of GnRHa upon alpha-subunit mRNA elevation and that differential cross-talk exists between the messengers.

Full text

PDF
329

Selected References

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

  1. Attardi B., Keeping H. S., Winters S. J., Kotsuji F., Troen P. Effect of inhibin from primate Sertoli cells and GnRH on gonadotropin subunit mRNA in rat pituitary cell cultures. Mol Endocrinol. 1989 Aug;3(8):1236–1242. doi: 10.1210/mend-3-8-1236. [DOI] [PubMed] [Google Scholar]
  2. Belchetz P. E., Plant T. M., Nakai Y., Keogh E. J., Knobil E. Hypophysial responses to continuous and intermittent delivery of hypopthalamic gonadotropin-releasing hormone. Science. 1978 Nov 10;202(4368):631–633. doi: 10.1126/science.100883. [DOI] [PubMed] [Google Scholar]
  3. Ben-Menahem D., Naor Z. Regulation of gonadotropin mRNA levels in cultured rat pituitary cells by gonadotropin-releasing hormone (GnRH): role for Ca2+ and protein kinase C. Biochemistry. 1994 Mar 29;33(12):3698–3704. doi: 10.1021/bi00178a029. [DOI] [PubMed] [Google Scholar]
  4. Ben-Menahem D., Shraga Z., Lewy H., Limor R., Hammel I., Stein R., Naor Z. Dissociation between release and gene expression of gonadotropin alpha-subunit in gonadotropin-releasing hormone-stimulated alpha T3-1 cell line. Biochemistry. 1992 Dec 29;31(51):12893–12898. doi: 10.1021/bi00166a026. [DOI] [PubMed] [Google Scholar]
  5. Cathala G., Savouret J. F., Mendez B., West B. L., Karin M., Martial J. A., Baxter J. D. A method for isolation of intact, translationally active ribonucleic acid. DNA. 1983;2(4):329–335. doi: 10.1089/dna.1983.2.329. [DOI] [PubMed] [Google Scholar]
  6. Chedrese P. J., Kay T. W., Jameson J. L. Gonadotropin-releasing hormone stimulates glycoprotein hormone alpha-subunit messenger ribonucleic acid (mRNA) levels in alpha T3 cells by increasing transcription and mRNA stability. Endocrinology. 1994 Jun;134(6):2475–2481. doi: 10.1210/endo.134.6.7515001. [DOI] [PubMed] [Google Scholar]
  7. Dan-Cohen H., Naor Z. Mechanism of action of gonadotropin releasing hormone upon gonadotropin secretion: involvement of protein kinase C as revealed by staurosporine inhibition and enzyme depletion. Mol Cell Endocrinol. 1990 Mar 5;69(2-3):135–144. doi: 10.1016/0303-7207(90)90007-u. [DOI] [PubMed] [Google Scholar]
  8. Gharib S. D., Wierman M. E., Shupnik M. A., Chin W. W. Molecular biology of the pituitary gonadotropins. Endocr Rev. 1990 Feb;11(1):177–199. doi: 10.1210/edrv-11-1-177. [DOI] [PubMed] [Google Scholar]
  9. Gordon D. F., Wood W. M., Ridgway E. C. Organization and nucleotide sequence of the mouse alpha-subunit gene of the pituitary glycoprotein hormones. DNA. 1988 Dec;7(10):679–690. doi: 10.1089/dna.1988.7.679. [DOI] [PubMed] [Google Scholar]
  10. Hoeffler J. P., Deutsch P. J., Lin J., Habener J. F. Distinct adenosine 3',5'-monophosphate and phorbol ester-responsive signal transduction pathways converge at the level of transcriptional activation by the interactions of DNA-binding proteins. Mol Endocrinol. 1989 May;3(5):868–880. doi: 10.1210/mend-3-5-868. [DOI] [PubMed] [Google Scholar]
  11. Horn F., Bilezikjian L. M., Perrin M. H., Bosma M. M., Windle J. J., Huber K. S., Blount A. L., Hille B., Vale W., Mellon P. L. Intracellular responses to gonadotropin-releasing hormone in a clonal cell line of the gonadotrope lineage. Mol Endocrinol. 1991 Mar;5(3):347–355. doi: 10.1210/mend-5-3-347. [DOI] [PubMed] [Google Scholar]
  12. Hubert J. F., Simard J., Gagné B., Barden N., Labrie F. Effect of luteinizing hormone releasing hormone (LHRH) and [D-Trp6, des-Gly-NH2(10)]LHRH ethylamide on alpha-subunit and LH beta messenger ribonucleic acid levels in rat anterior pituitary cells in culture. Mol Endocrinol. 1988 Jun;2(6):521–527. doi: 10.1210/mend-2-6-521. [DOI] [PubMed] [Google Scholar]
  13. Kay T. W., Jameson J. L. Identification of a gonadotropin-releasing hormone-responsive region in the glycoprotein hormone alpha-subunit promoter. Mol Endocrinol. 1992 Nov;6(11):1767–1773. doi: 10.1210/mend.6.11.1282668. [DOI] [PubMed] [Google Scholar]
  14. Kim M. K., McClaskey J. H., Bodenner D. L., Weintraub B. D. An AP-1-like factor and the pituitary-specific factor Pit-1 are both necessary to mediate hormonal induction of human thyrotropin beta gene expression. J Biol Chem. 1993 Nov 5;268(31):23366–23375. [PubMed] [Google Scholar]
  15. Liu F., Thompson M. A., Wagner S., Greenberg M. E., Green M. R. Activating transcription factor-1 can mediate Ca(2+)- and cAMP-inducible transcriptional activation. J Biol Chem. 1993 Mar 25;268(9):6714–6720. [PubMed] [Google Scholar]
  16. Lucas P. C., Granner D. K. Hormone response domains in gene transcription. Annu Rev Biochem. 1992;61:1131–1173. doi: 10.1146/annurev.bi.61.070192.005411. [DOI] [PubMed] [Google Scholar]
  17. Mercer J. E. Pituitary gonadotropin gene regulation. Mol Cell Endocrinol. 1990 Oct 1;73(1):C63–C67. doi: 10.1016/0303-7207(90)90037-9. [DOI] [PubMed] [Google Scholar]
  18. Miner J. N., Yamamoto K. R. Regulatory crosstalk at composite response elements. Trends Biochem Sci. 1991 Nov;16(11):423–426. doi: 10.1016/0968-0004(91)90168-u. [DOI] [PubMed] [Google Scholar]
  19. Mischak H., Goodnight J. A., Kolch W., Martiny-Baron G., Schaechtle C., Kazanietz M. G., Blumberg P. M., Pierce J. H., Mushinski J. F. Overexpression of protein kinase C-delta and -epsilon in NIH 3T3 cells induces opposite effects on growth, morphology, anchorage dependence, and tumorigenicity. J Biol Chem. 1993 Mar 25;268(9):6090–6096. [PubMed] [Google Scholar]
  20. Naor Z., Capponi A. M., Rossier M. F., Ayalon D., Limor R. Gonadotropin-releasing hormone-induced rise in cytosolic free Ca2+ levels: mobilization of cellular and extracellular Ca2+ pools and relationship to gonadotropin secretion. Mol Endocrinol. 1988 Jun;2(6):512–520. doi: 10.1210/mend-2-6-512. [DOI] [PubMed] [Google Scholar]
  21. Naor Z., Dan-Cohen H., Hermon J., Limor R. Induction of exocytosis in permeabilized pituitary cells by alpha- and beta-type protein kinase C. Proc Natl Acad Sci U S A. 1989 Jun;86(12):4501–4504. doi: 10.1073/pnas.86.12.4501. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Naor Z., Koch Y., Chobsieng P., Zor U. Pituitary cyclic AMP production and mechanism of luteinizing hormone release. FEBS Lett. 1975 Oct 15;58(1):318–321. doi: 10.1016/0014-5793(75)80288-2. [DOI] [PubMed] [Google Scholar]
  23. Naor Z., Shearman M. S., Kishimoto A., Nishizuka Y. Calcium-independent activation of hypothalamic type I protein kinase C by unsaturated fatty acids. Mol Endocrinol. 1988 Nov;2(11):1043–1048. doi: 10.1210/mend-2-11-1043. [DOI] [PubMed] [Google Scholar]
  24. Naor Z. Signal transduction mechanisms of Ca2+ mobilizing hormones: the case of gonadotropin-releasing hormone. Endocr Rev. 1990 May;11(2):326–353. doi: 10.1210/edrv-11-2-326. [DOI] [PubMed] [Google Scholar]
  25. Nishizuka Y. Intracellular signaling by hydrolysis of phospholipids and activation of protein kinase C. Science. 1992 Oct 23;258(5082):607–614. doi: 10.1126/science.1411571. [DOI] [PubMed] [Google Scholar]
  26. Pierce J. G., Parsons T. F. Glycoprotein hormones: structure and function. Annu Rev Biochem. 1981;50:465–495. doi: 10.1146/annurev.bi.50.070181.002341. [DOI] [PubMed] [Google Scholar]
  27. Rüegg U. T., Burgess G. M. Staurosporine, K-252 and UCN-01: potent but nonspecific inhibitors of protein kinases. Trends Pharmacol Sci. 1989 Jun;10(6):218–220. doi: 10.1016/0165-6147(89)90263-0. [DOI] [PubMed] [Google Scholar]
  28. Salton S. R., Blum M., Jonassen J. A., Clayton R. N., Roberts J. L. Stimulation of pituitary luteinizing hormone secretion by gonadotropin-releasing hormone is not coupled to beta-luteinizing hormone gene transcription. Mol Endocrinol. 1988 Nov;2(11):1033–1042. doi: 10.1210/mend-2-11-1033. [DOI] [PubMed] [Google Scholar]
  29. Schoderbek W. E., Roberson M. S., Maurer R. A. Two different DNA elements mediate gonadotropin releasing hormone effects on expression of the glycoprotein hormone alpha-subunit gene. J Biol Chem. 1993 Feb 25;268(6):3903–3910. [PubMed] [Google Scholar]
  30. Shupnik M. A. Effects of gonadotropin-releasing hormone on rat gonadotropin gene transcription in vitro: requirement for pulsatile administration for luteinizing hormone-beta gene stimulation. Mol Endocrinol. 1990 Oct;4(10):1444–1450. doi: 10.1210/mend-4-10-1444. [DOI] [PubMed] [Google Scholar]
  31. Starzec A., Jutisz M., Counis R. Cyclic adenosine monophosphate and phorbol ester, like gonadotropin-releasing hormone, stimulate the biosynthesis of luteinizing hormone polypeptide chains in a nonadditive manner. Mol Endocrinol. 1989 Apr;3(4):618–624. doi: 10.1210/mend-3-4-618. [DOI] [PubMed] [Google Scholar]
  32. Stojilković S. S., Catt K. J. Calcium oscillations in anterior pituitary cells. Endocr Rev. 1992 May;13(2):256–280. doi: 10.1210/edrv-13-2-256. [DOI] [PubMed] [Google Scholar]
  33. Toullec D., Pianetti P., Coste H., Bellevergue P., Grand-Perret T., Ajakane M., Baudet V., Boissin P., Boursier E., Loriolle F. The bisindolylmaleimide GF 109203X is a potent and selective inhibitor of protein kinase C. J Biol Chem. 1991 Aug 25;266(24):15771–15781. [PubMed] [Google Scholar]
  34. Tsien R. Y. New calcium indicators and buffers with high selectivity against magnesium and protons: design, synthesis, and properties of prototype structures. Biochemistry. 1980 May 27;19(11):2396–2404. doi: 10.1021/bi00552a018. [DOI] [PubMed] [Google Scholar]
  35. Weiss J., Jameson J. L., Burrin J. M., Crowley W. F., Jr Divergent responses of gonadotropin subunit messenger RNAs to continuous versus pulsatile gonadotropin-releasing hormone in vitro. Mol Endocrinol. 1990 Apr;4(4):557–564. doi: 10.1210/mend-4-4-557. [DOI] [PubMed] [Google Scholar]
  36. Windle J. J., Weiner R. I., Mellon P. L. Cell lines of the pituitary gonadotrope lineage derived by targeted oncogenesis in transgenic mice. Mol Endocrinol. 1990 Apr;4(4):597–603. doi: 10.1210/mend-4-4-597. [DOI] [PubMed] [Google Scholar]
  37. Wondisford F. E., Steinfelder H. J., Nations M., Radovick S. AP-1 antagonizes thyroid hormone receptor action on the thyrotropin beta-subunit gene. J Biol Chem. 1993 Feb 5;268(4):2749–2754. [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES