Stimulation of mitogen-activated protein kinase by gonadotropin-releasing hormone in human granulosa-luteal cells

Keun Kang Sung Keun Kang, C. J. Tai, P. S. Nathwani, K. C. Choi, P. C K Leung

Research output: Contribution to journalArticle

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Abstract

The present study investigated the activation of mitogen activated protein kinases (MAPKs) by a GnRH agonist (GnRHa) in human granulosa-luteal cells (hGLCs). The phosphorylation state of p44 and p42 MAPK was examined using antibodies that distinguish phosphop44/42 MAPK (Thr202/Tyr204) from total p44/42 MAPK (activated plus inactivated). Activation of MAPK by GnRHa was observed within 5 min and was sustained for 60 min after treatment. GnRHa stimulated MAPK activation in a dose-dependent manner, with maximum stimulation (6.7-fold over basal levels) at 10-7 M. Pretreatment with a protein kinase C (PKC) inhibitor, GF109203X, completely blocked GnRHa-induced MAPK activation. In addition, pretreatment with a PKC activator, phorbol-12-myristate 13-acetate, potentiated GnRH-induced MAPK activation. These results indicate that GnRHa stimulates MAPK activation through a PKC-dependent pathway in hGLCs, possibly coupled to Gqα protein. MAPK activation was also observed in response to 8-bromo-cAMP or cholera toxin, but not pertussis toxin. Forskolin (50 μM) substantially stimulated a rapid cAMP accumulation, whereas GnRHa (10-7 M) or pertussis toxin (100 mg/ml) did not affect basal intracellular cAMP levels. Cotreatment of GnRHa (10-7 M) did not attenuate forskolin- or hCG-stimulated cAMP accumulation. These results suggest that the GnRH receptor is probably not coupled to Gsα or Giα in hGLCs. Finally, GnRHa (10-7 M) stimulated a significant increase in Elk-1 phosphorylation and c-fos messenger RNA expression, as revealed by an in vitro kinase assay and Northern blot analysis, respectively. These results clearly demonstrate that GnRH activates the MAPK cascade through a PKC-dependent pathway in the human ovary.

Original languageEnglish
Pages (from-to)671-679
Number of pages9
JournalEndocrinology
Volume142
Issue number2
DOIs
Publication statusPublished - 2001
Externally publishedYes

Fingerprint

Luteal Cells
Mitogen-Activated Protein Kinases
Gonadotropin-Releasing Hormone
Protein Kinase C
Pertussis Toxin
Colforsin
Gq-G11 GTP-Binding Protein alpha Subunits
Phosphorylation
LHRH Receptors
8-Bromo Cyclic Adenosine Monophosphate
Protein C Inhibitor
Cholera Toxin
Mitogen-Activated Protein Kinase 1
Protein Kinase Inhibitors
Northern Blotting
Ovary
Acetates
Phosphotransferases

ASJC Scopus subject areas

  • Endocrinology
  • Endocrinology, Diabetes and Metabolism

Cite this

Stimulation of mitogen-activated protein kinase by gonadotropin-releasing hormone in human granulosa-luteal cells. / Sung Keun Kang, Keun Kang; Tai, C. J.; Nathwani, P. S.; Choi, K. C.; Leung, P. C K.

In: Endocrinology, Vol. 142, No. 2, 2001, p. 671-679.

Research output: Contribution to journalArticle

Sung Keun Kang, Keun Kang ; Tai, C. J. ; Nathwani, P. S. ; Choi, K. C. ; Leung, P. C K. / Stimulation of mitogen-activated protein kinase by gonadotropin-releasing hormone in human granulosa-luteal cells. In: Endocrinology. 2001 ; Vol. 142, No. 2. pp. 671-679.
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