title: WNT5a is required for normal ovarian follicle development and antagonizes gonadotropin responsiveness in granulosa cells by suppressing canonical WNT signaling.
abstract: Whereas the roles of the canonical wingless-type MMTV (mouse mammary tumor virus) integration site family (WNT) signaling pathway in the regulation of ovarian follicle growth and steroidogenesis are now established, noncanonical WNT signaling in the ovary has been largely overlooked. Noncanonical WNTs, including WNT5a and WNT11, are expressed in granulosa cells (GCs) and are differentially regulated throughout follicle development, but their physiologic roles remain unknown. Using conditional gene targeting, we found that GC-specific inactivation ofWnt5a(but notWnt11) results in the female subfertility associated with increased follicular atresia and decreased rates of ovulation. Microarray analyses have revealed that WNT5a acts to down-regulate the expression of FSH-responsive genesin vitro, and corresponding increases in the expression of these genes have been found in the GCs of conditional knockout mice. Unexpectedly, we found that WNT5a regulates its target genes not by signalingviathe WNT/Ca(2+)or planar cell polarity pathways, but rather by inhibiting the canonical pathway, causing both β-catenin (CTNNB1) and cAMP responsive element binding (CREB) protein levels to decreaseviaa glycogen synthase kinase-3β-dependent mechanism. We further found that WNT5a prevents follicle-stimulating hormone and luteinizing protein from up-regulating the CTNNB1 and CREB proteins and their target genes, indicating that WNT5a functions as a physiologic inhibitor of gonadotropin signaling. Together, these findings identify WNT5a as a key regulator of follicle development and gonadotropin responsiveness.-Abedini, A., Zamberlam, G., Lapointe, E., Tourigny, C., Boyer, A., Paquet, M., Hayashi, K., Honda, H., Kikuchi, A., Price, C., Boerboom, D. WNT5a is required for normal ovarian follicle development and antagonizes gonadotropin responsiveness in granulosa cells by suppressing canonical WNT signaling.
authors.affiliation: *Centre de Recherche en Reproduction Animale, Faculté de Médecine Vétérinaire, Université de Montréal, Saint-Hyacinthe, Québec, Canada; Department of Physiology, Southern Illinois University School of Medicine, Carbondale, Illinois, USA; Department of Disease Model, Research Institute for Radiation Biology and Medicine, Hiroshima University, Hiroshima, Japan; and Department of Molecular Biology and Biochemistry, Graduate School of Medicine, Osaka University, Osaka, Japan.
authors.forename: Atefeh
authors.initials: A
authors.lastname: Abedini
authors.affiliation: *Centre de Recherche en Reproduction Animale, Faculté de Médecine Vétérinaire, Université de Montréal, Saint-Hyacinthe, Québec, Canada; Department of Physiology, Southern Illinois University School of Medicine, Carbondale, Illinois, USA; Department of Disease Model, Research Institute for Radiation Biology and Medicine, Hiroshima University, Hiroshima, Japan; and Department of Molecular Biology and Biochemistry, Graduate School of Medicine, Osaka University, Osaka, Japan.
authors.forename: Gustavo
authors.initials: G
authors.lastname: Zamberlam
authors.affiliation: *Centre de Recherche en Reproduction Animale, Faculté de Médecine Vétérinaire, Université de Montréal, Saint-Hyacinthe, Québec, Canada; Department of Physiology, Southern Illinois University School of Medicine, Carbondale, Illinois, USA; Department of Disease Model, Research Institute for Radiation Biology and Medicine, Hiroshima University, Hiroshima, Japan; and Department of Molecular Biology and Biochemistry, Graduate School of Medicine, Osaka University, Osaka, Japan.
authors.forename: Evelyne
authors.initials: E
authors.lastname: Lapointe
authors.affiliation: *Centre de Recherche en Reproduction Animale, Faculté de Médecine Vétérinaire, Université de Montréal, Saint-Hyacinthe, Québec, Canada; Department of Physiology, Southern Illinois University School of Medicine, Carbondale, Illinois, USA; Department of Disease Model, Research Institute for Radiation Biology and Medicine, Hiroshima University, Hiroshima, Japan; and Department of Molecular Biology and Biochemistry, Graduate School of Medicine, Osaka University, Osaka, Japan.
authors.forename: Catherine
authors.initials: C
authors.lastname: Tourigny
authors.affiliation: *Centre de Recherche en Reproduction Animale, Faculté de Médecine Vétérinaire, Université de Montréal, Saint-Hyacinthe, Québec, Canada; Department of Physiology, Southern Illinois University School of Medicine, Carbondale, Illinois, USA; Department of Disease Model, Research Institute for Radiation Biology and Medicine, Hiroshima University, Hiroshima, Japan; and Department of Molecular Biology and Biochemistry, Graduate School of Medicine, Osaka University, Osaka, Japan.
authors.forename: Alexandre
authors.initials: A
authors.lastname: Boyer
authors.affiliation: *Centre de Recherche en Reproduction Animale, Faculté de Médecine Vétérinaire, Université de Montréal, Saint-Hyacinthe, Québec, Canada; Department of Physiology, Southern Illinois University School of Medicine, Carbondale, Illinois, USA; Department of Disease Model, Research Institute for Radiation Biology and Medicine, Hiroshima University, Hiroshima, Japan; and Department of Molecular Biology and Biochemistry, Graduate School of Medicine, Osaka University, Osaka, Japan.
authors.forename: Marilène
authors.initials: M
authors.lastname: Paquet
authors.affiliation: *Centre de Recherche en Reproduction Animale, Faculté de Médecine Vétérinaire, Université de Montréal, Saint-Hyacinthe, Québec, Canada; Department of Physiology, Southern Illinois University School of Medicine, Carbondale, Illinois, USA; Department of Disease Model, Research Institute for Radiation Biology and Medicine, Hiroshima University, Hiroshima, Japan; and Department of Molecular Biology and Biochemistry, Graduate School of Medicine, Osaka University, Osaka, Japan.
authors.forename: Kanako
authors.initials: K
authors.lastname: Hayashi
authors.affiliation: *Centre de Recherche en Reproduction Animale, Faculté de Médecine Vétérinaire, Université de Montréal, Saint-Hyacinthe, Québec, Canada; Department of Physiology, Southern Illinois University School of Medicine, Carbondale, Illinois, USA; Department of Disease Model, Research Institute for Radiation Biology and Medicine, Hiroshima University, Hiroshima, Japan; and Department of Molecular Biology and Biochemistry, Graduate School of Medicine, Osaka University, Osaka, Japan.
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Honda
authors.affiliation: *Centre de Recherche en Reproduction Animale, Faculté de Médecine Vétérinaire, Université de Montréal, Saint-Hyacinthe, Québec, Canada; Department of Physiology, Southern Illinois University School of Medicine, Carbondale, Illinois, USA; Department of Disease Model, Research Institute for Radiation Biology and Medicine, Hiroshima University, Hiroshima, Japan; and Department of Molecular Biology and Biochemistry, Graduate School of Medicine, Osaka University, Osaka, Japan.
authors.forename: Akira
authors.initials: A
authors.lastname: Kikuchi
authors.affiliation: *Centre de Recherche en Reproduction Animale, Faculté de Médecine Vétérinaire, Université de Montréal, Saint-Hyacinthe, Québec, Canada; Department of Physiology, Southern Illinois University School of Medicine, Carbondale, Illinois, USA; Department of Disease Model, Research Institute for Radiation Biology and Medicine, Hiroshima University, Hiroshima, Japan; and Department of Molecular Biology and Biochemistry, Graduate School of Medicine, Osaka University, Osaka, Japan.
authors.forename: Christopher
authors.initials: C
authors.lastname: Price
authors.affiliation: *Centre de Recherche en Reproduction Animale, Faculté de Médecine Vétérinaire, Université de Montréal, Saint-Hyacinthe, Québec, Canada; Department of Physiology, Southern Illinois University School of Medicine, Carbondale, Illinois, USA; Department of Disease Model, Research Institute for Radiation Biology and Medicine, Hiroshima University, Hiroshima, Japan; and Department of Molecular Biology and Biochemistry, Graduate School of Medicine, Osaka University, Osaka, Japan derek.boerboom@umontreal.ca.
authors.forename: Derek
authors.initials: D
authors.lastname: Boerboom
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D017362:Cyclic AMP Response Element-Binding Protein / Q000235:genetics / Q000378:metabolism; D005260:Female; D006062:Gonadotropins / Q000494:pharmacology*; D006107:Granulosa Cells / Q000187:drug effects* / Q000378:metabolism; D015151:Immunoblotting; D007150:Immunohistochemistry; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D008822:Mice, Transgenic; D020411:Oligonucleotide Array Sequence Analysis; D006080:Ovarian Follicle / Q000166:cytology / Q000254:growth & development / Q000378:metabolism*; D010060:Ovulation / Q000187:drug effects / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D059467:Transcriptome / Q000187:drug effects / Q000235:genetics; D051153:Wnt Proteins / Q000235:genetics* / Q000378:metabolism; D060449:Wnt Signaling Pathway / Q000187:drug effects* / Q000235:genetics; D000071818:Wnt-5a Protein
chemical_list: C492004:Creb1 protein, mouse; D017362:Cyclic AMP Response Element-Binding Protein; D006062:Gonadotropins; D051153:Wnt Proteins; D000071818:Wnt-5a Protein; C438615:Wnt5a protein, mouse
keywords: conditional gene targeting; female fertility; microarray