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J Biol Chem, Vol. 275, Issue 13, 9193-9200, March 31, 2000

Gonadotropin-releasing Hormone Receptor Initiates Multiple Signaling Pathways by Exclusively Coupling to Gq/11 Proteins*

Robert Grosse, Andrea Schmid, Torsten Schöneberg, Andreas Herrlich, Peter MuhnDagger , Günter Schultz, and Thomas Gudermann§

From the Institut für Pharmakologie, Freie Universität Berlin, Thielallee 69-73, D-14195 Berlin, Germany and the Dagger  Forschungslaboratorien der Schering AG, 13342 Berlin, Germany

The agonist-bound gonadotropin-releasing hormone (GnRH) receptor engages several distinct signaling cascades, and it has recently been proposed that coupling of a single type of receptor to multiple G proteins (Gq, Gs, and Gi) is responsible for this behavior. GnRH-dependent signaling was studied in gonadotropic alpha T3-1 cells endogenously expressing the murine receptor and in CHO-K1 (CHO#3) and COS-7 cells transfected with the human GnRH receptor cDNA. In all cell systems studied, GnRH-induced phospholipase C activation and Ca2+ mobilization was pertussis toxin-insensitive, as was GnRH-mediated extracellular signal-regulated kinase activation. Whereas the Gi-coupled m2 muscarinic receptor interacted with a chimeric Gs protein (Gsi5) containing the C-terminal five amino acids of Galpha i2, the human GnRH receptor was unable to activate the G protein chimera. GnRH challenge of alpha T3-1, CHO#3 and of GnRH receptor-expressing COS-7 cells did not result in agonist-dependent cAMP formation. GnRH challenge of CHO#3 cells expressing a cAMP-responsive element-driven firefly luciferase did not result in increased reporter gene expression. However, coexpression of the human GnRH receptor and adenylyl cyclase I in COS-7 cells led to clearly discernible GnRH-dependent cAMP formation subsequent to GnRH-elicited rises in [Ca2+]i. In alpha T3-1 and CHO#3 cell membranes, addition of [alpha -32P]GTP azidoanilide resulted in GnRH receptor-dependent labeling of Galpha q/11 but not of Galpha i, Galpha s or Galpha 12/13 proteins. Thus, the murine and human GnRH receptors exclusively couple to G proteins of the Gq/11 family. Multiple GnRH-dependent signaling pathways are therefore initiated downstream of the receptor/G protein interface and are not indicative of a multiple G protein coupling potential of the GnRH receptor.

* This work was supported by the Deutsche Forschungsgemeinschaft and Fonds der Chemischen Industrie.The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

§ To whom correspondence should be addressed. Tel.: 49-30-8445-1818; Fax: 49-30-162-8445-1818; E-mail: guderman@zedat.fu-berlin.de.

Copyright © 2000 by The American Society for Biochemistry and Molecular Biology, Inc.
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