title: Fibroblast growth factors activate mitogen-activated protein kinase pathways to promote migration in ovine trophoblast cells.
abstract: Fibroblast growth factors (FGFs) 2 and FGF10 are uterine- and conceptus-derived factors that mediate trophoblast activities in cattle and sheep. To extend our understanding of how FGFs may control peri-implantation development in ruminants, we determined whether FGF2 and FGF10 impact trophoblast cell migration. Transwell inserts containing 8 μm pores were used to examine whether FGF2 or FGF10 supplementation increased oTr1 cell migration. Supplementation with 0.5 ng/ml FGF2 or FGF10 did not affect oTr1 cell migration number, but exposure to 5 or 50 ng/ml FGF2 or FGF10 increased (P<0.05) oTr1 cell migration when compared with controls. The involvement of specific MAP kinase (MAPK) cascades in mediating this FGF response was examined by using pharmacological inhibitors of specific MAPKs. Western blot analysis indicated that FGF2 and FGF10 increased phosphorylation status of MAPKs 1, 3, 8, 9, and 14. Exposure to specific inhibitors blocked FGF induction of each MAPK. Exposure to inhibitors before supplementation with FGF2 or FGF10 prevented FGF induction of cell migration, indicating that each of these signaling molecules was required for FGF effects. A final series of studies examined whether FGF2 and FGF10 also mediated the migration of a bovine trophoblast line (CT1 cell). Increases in migration were detected in each cell line by supplementing 5 or 50 ng/ml FGF2 or FGF10 (P<0.05). In summary, FGF2 and FGF10 regulate migratory activity of ovine trophoblast cells through MAPK-dependent pathways. These outcomes provide further evidence that FGFs function as mediators of peri-implantation conceptus development in cattle and sheep.
authors.affiliation: Department of Animal Sciences, DH Barron Reproductive and Perinatal Biology Research Program, University of Florida, PO Box 110910, Gainesville, Florida 32611-0910, USA.
authors.forename: Qi En
authors.initials: QE
authors.lastname: Yang
authors.forename: Mariana I
authors.initials: MI
authors.lastname: Giassetti
authors.forename: Alan D
authors.initials: AD
authors.lastname: Ealy
mesh_terms: D000704:Analysis of Variance; D000818:Animals; D015153:Blotting, Western; D002417:Cattle; D002460:Cell Line; D054443:Cell Migration Assays; D002465:Cell Movement / Q000187:drug effects; D004789:Enzyme Activation; D005260:Female; D051526:Fibroblast Growth Factor 10 / Q000378:metabolism*; D016222:Fibroblast Growth Factor 2 / Q000378:metabolism*; D006801:Humans; D020935:MAP Kinase Signaling System / Q000187:drug effects; D020928:Mitogen-Activated Protein Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D010766:Phosphorylation; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D011994:Recombinant Proteins / Q000378:metabolism; D012756:Sheep; D013997:Time Factors; D014327:Trophoblasts / Q000187:drug effects / Q000201:enzymology*
chemical_list: C496463:FGF10 protein, human; D051526:Fibroblast Growth Factor 10; D047428:Protein Kinase Inhibitors; D011994:Recombinant Proteins; D016222:Fibroblast Growth Factor 2; D020928:Mitogen-Activated Protein Kinases