FGF-2 alters the fate of mouse epiblast from ectoderm to mesoderm in vitro.
We have developed an in vitro differentiation assay to characterize the ability of peptide growth factors to induce differentiation in mouse epiblast. We report that culturing explants of mouse anterior epiblast, a tissue normally fated to give rise to neuroectoderm and surface ectoderm, in a serum-free, chemically defined medium with 10-50 ng/ml of FGF-2 induced gross changes in cell morphology. Treated cells adopted an elongated, flattened morphology but did not migrate from the explant. Instead, FGF-2-treated cells condensed into multicellular mounds or ridges. Immunocytochemistry showed that cells in treated explants expressed vimentin and in situ hybridization demonstrated that FGF-2 induced the expression of brachyury, goosecoid, and myo-D in regions of treated explants displaying morphological differentiation. Control explants cultured with platelet-derived growth factor AA (PDGF AA), transforming growth factor-beta 1 (TGF-beta 1), or in defined medium alone showed no morphological or biochemical differentiation. These results indicate that FGF-2 altered the fate of mouse anterior epiblast from ectoderm to mesoderm in vitro. Cell migration, which is characteristic of primitive streak mesoderm in vivo, was not induced by FGF-2 in these assays. However, the changes in morphology and the expression of mesodermal genes in vitro do support an early role for FGF signaling in the induction of mouse primitive streak mesoderm, as well as in later patterning events during embryogenesis.
Burdsal
C A
CA
Department Cell and Molecular Biology, Tulane University, New Orleans, Louisiana 70118, USA. cburdsal@mailhost.tcs.tulane.edu
Flannery
M L
ML
Pedersen
R A
RA
D000818:Animals; D004475:Ectoderm / Q000187:drug effects* / Q000502:physiology; D004627:Embryonic Induction; D005260:Female; D016222:Fibroblast Growth Factor 2 / Q000494:pharmacology*; D006801:Humans; D008648:Mesoderm / Q000502:physiology*; D051379:Mice; D008813:Mice, Inbred ICR; D017570:MyoD Protein / Q000032:analysis; D016133:Polymerase Chain Reaction; D011247:Pregnancy
D017570:MyoD Protein; D016222:Fibroblast Growth Factor 2