title: Signalling pathways involved in the synergistic effects of human growth differentiation factor 9 and bone morphogenetic protein 15.
abstract: Growth differentiation factor 9 (GDF9) and bone morphogenetic protein 15 (BMP15) act synergistically to regulate granulosa cell proliferation and steroid production in several species. Several non-Sma and mothers against decapentaplegic (SMAD) signalling pathways are involved in the action of murine and ovine GDF9 and BMP15 in combination, with the pathways utilised differing between the two species. The aims of this research were to determine if human GDF9 and BMP15 also act in a synergistic manner to stimulate granulosa cell proliferation and to identify which non-SMAD signalling pathways are activated. Human GDF9 with BMP15 (GDF9+BMP15) stimulated an increase in (3)H-thymidine incorporation (P<0.001), which was greater than the increase with BMP15 alone, while GDF9 alone had no effect. The stimulation of (3)H-thymidine incorporation by GDF9+BMP15 was reduced by the addition of inhibitors to the SMAD2/3, nuclear factor-KB (NF-KB) and c-Jun N-terminal kinase (JNK) signalling pathways. Inhibitors to the SMAD1/5/8, extracellular signal-regulated kinase mitogen-activated protein kinase (ERK-MAPK) or p38-MAPK pathways had no effect. The addition of the BMP receptor 2 (BMPR2) extracellular domain also inhibited stimulation of (3)H-thymidine incorporation by GDF9+BMP15. In conclusion, human GDF9 and BMP15 act synergistically to stimulate granulosa cell proliferation, a response that also involves species-specific non-SMAD signalling pathways.
authors.affiliation: AgResearch, Animal Productivity, Invermay Agricultural Centre, Private Bag 50034, Mosgiel 9053, New Zealand.
authors.forename: Karen L
authors.initials: KL
authors.lastname: Reader
authors.affiliation: Robinson Research Institute, School of Paediatrics and Reproductive Health, University of Adelaide, GPO Box 498, Adelaide, SA, Australia.
authors.forename: David G
authors.initials: DG
authors.lastname: Mottershead
authors.affiliation: Robinson Research Institute, School of Paediatrics and Reproductive Health, University of Adelaide, GPO Box 498, Adelaide, SA, Australia.
authors.forename: Georgia A
authors.initials: GA
authors.lastname: Martin
authors.affiliation: Robinson Research Institute, School of Paediatrics and Reproductive Health, University of Adelaide, GPO Box 498, Adelaide, SA, Australia.
authors.forename: Robert B
authors.initials: RB
authors.lastname: Gilchrist
authors.affiliation: Victoria University of Wellington, School of Biological Sciences, PO Box 600, Wellington, New Zealand.
authors.forename: Derek A
authors.initials: DA
authors.lastname: Heath
authors.affiliation: Victoria University of Wellington, School of Biological Sciences, PO Box 600, Wellington, New Zealand.
authors.forename: Kenneth P
authors.initials: KP
authors.lastname: McNatty
authors.affiliation: AgResearch, Animal Productivity, Invermay Agricultural Centre, Private Bag 50034, Mosgiel 9053, New Zealand.
authors.forename: Jennifer L
authors.initials: JL
authors.lastname: Juengel
mesh_terms: D000818:Animals; D055430:Bone Morphogenetic Protein 15 / Q000494:pharmacology*; D049109:Cell Proliferation / Q000187:drug effects*; D004305:Dose-Response Relationship, Drug; D004357:Drug Synergism; D005260:Female; D006107:Granulosa Cells / Q000187:drug effects* / Q000378:metabolism; D055429:Growth Differentiation Factor 9 / Q000494:pharmacology*; D006801:Humans; D048031:JNK Mitogen-Activated Protein Kinases / Q000378:metabolism; D057507:Mice, 129 Strain; D016328:NF-kappa B / Q000378:metabolism; D017207:Rats, Sprague-Dawley; D015290:Second Messenger Systems / Q000187:drug effects*; D051896:Smad Proteins, Receptor-Regulated / Q000378:metabolism
chemical_list: C525901:BMP15 protein, human; D055430:Bone Morphogenetic Protein 15; C525897:GDF9 protein, human; D055429:Growth Differentiation Factor 9; D016328:NF-kappa B; D051896:Smad Proteins, Receptor-Regulated; D048031:JNK Mitogen-Activated Protein Kinases