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J. Biol. Chem., Vol. 275, Issue 36, 27615-27626, September 8, 2000
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From the Follicle-stimulating hormone (FSH) regulated
growth and function of the ovarian follicle was previously thought to
be mediated solely through activation of Gs-coupled
receptors. In this study, we show for the first time that this function
is predominantly mediated through the alternatively spliced and novel
growth factor type 1 receptor (oFSH-R3) that is also present in the
ovary. Immortalized granulosa cells lacking endogenous FSH receptors,
when transfected with either oFSH-R3 cDNA (JC-R3) or the
Gs-coupled oFSH-R1 (JC-R1), expressed the corresponding
glycosylated receptor. In JC-R3 or JC-R1 cells labeled with
bromodeoxyuridine or [3H]thymidine, FSH stimulated
the cells to progress through S-phase and divide. The growth promoting
effect of recombinant FSH in JC-R3 cells was preceded by the rapid
activation of ERK1 and ERK2. This effect was hormone-specific and
transient. In JC-R3 cells inhibitors like calphostin C,
PD98059, Ag 18, or calcium chelators EGTA or
1,2-bis(O-aminophenoxy)ethane-N,N,N',N'-tetraacetic
acid/AM inhibited both mitogen-activated protein kinase activation and bromodeoxyuridine incorporation. FSH induced phosphorylation of the
FSH-R3 receptor was blocked by pretreating cells with calphostin C. There was no cAMP induction by FSH in JC-R3 cells. The cAMP independent
growth promoting effect of FSH is mediated by activation of
Ca2+ and mitogen-activated protein
kinase-dependent pathways. Thus, alternative splicing of a
G-protein coupled receptor creates the expression of a novel receptor
motif that can mediate a widely recognized function of the glycoprotein hormone.
Activation of Extracellular-regulated Kinase Pathways in
Ovarian Granulosa Cells by the Novel Growth Factor Type 1 Follicle-stimulating Hormone Receptor
ROLE IN HORMONE SIGNALING AND CELL PROLIFERATION*
,,¶
Molecular Reproduction Research Laboratory,
Clinical Research Institute of Montreal, Montreal, Québec H2W
1R7, Canada and the § Department of Obstetrics, Gynecology
and Reproductive Sciences, University of Saskatchewan,
Saskatoon S7N 0W8, Canada
*
This work was supported by a grant from the Medical Research
Council of Canada.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.
Copyright © 2000 by The American Society for Biochemistry and Molecular Biology, Inc.
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