Dependence of GnRH-induced phosphorylation of CREB and BAD on EGF receptor transactivation in GT1-7 neuronal cells.
The hypothalamic neuropeptide, gonadotropin releasing hormone (GnRH), is a primary regulatory factor in the neuroendocrine control of reproduction. The GnRH decapeptide is released in an episodic manner from hypothalamic GnRH neurons, which are known to express GnRH receptors. Here we examined the signaling pathways by which autocrine GnRH stimulation generates cell survival and proliferative signals in hypothalamic GT1-7 cells. Both GnRH and epidermal growth factor (EGF) caused rapid phosphorylation of cyclic AMP response element binding protein (CREB) and BAD. The selective epidermal growth factor receptor (EGF-R) antagonist, AG1478, attenuates the phosphorylation of these proteins by GnRH and EGF. Inhibition of PKC and Src abolished the stimulatory effects of GnRH, but not that of EGF, consistent with a critical role of these signaling molecules upstream of the EGF-R. All of these effects of GnRH were mimicked by phorbol 12 myristate 13-acetate (PMA). Consistent with the prosurvival and mitogenic effects of phosphoinositide 3-kinase/Akt (P13-K/Akt) downstream of the EGF-R, inhibition of P13-K diminished the activation of these proteins following stimulation with GnRH, EGF, and PMA. Overexpression of dominant negative Akt attenuated agonist-induced phosphorylation of BAD, but not that of ERK1/2 and CREB. Moreover, overexpression of wild-type RSK-1 resulted in enhanced basal as well as agonist-induced phosphorylation of CREB and BAD, indicating a critical role of RSK-1 in activating cytosolic as well as nuclear proteins. These data reveal novel signaling mechanisms of GnRH-induced phosphorylation of CREB and BAD in GT1-7 neurons through transactivation of the EGF-R.
Neithardt
Adrienne
A
Section on Hormonal Regulation, Endocrinology and Reproduction Research Branch, NICHD, National Institutes of Health, Bethesda, Maryland 20892-4510, USA.
Farshori
M Parvaiz
MP
Shah
Farzana B
FB
Catt
Kevin J
KJ
Shah
Bukhtiar H
BH
D000818:Animals; D002460:Cell Line; D017362:Cyclic AMP Response Element-Binding Protein / Q000378:metabolism*; D004815:Epidermal Growth Factor / Q000494:pharmacology; D066246:ErbB Receptors / Q000187:drug effects / Q000502:physiology*; D007987:Gonadotropin-Releasing Hormone / Q000494:pharmacology*; D007031:Hypothalamus / Q000502:physiology; D051379:Mice; D009474:Neurons / Q000502:physiology; D010766:Phosphorylation; D051025:bcl-Associated Death Protein / Q000378:metabolism*
D017362:Cyclic AMP Response Element-Binding Protein; D051025:bcl-Associated Death Protein; D007987:Gonadotropin-Releasing Hormone; D004815:Epidermal Growth Factor; D066246:ErbB Receptors