Transplacental delivery of the Wnt antagonist Frzb1 inhibits development of caudal paraxial mesoderm and skeletal myogenesis in mouse embryos.
Axial structures (neural tube/notochord) and surface ectoderm activate myogenesis in the mouse embryo; their action can be reproduced, at least in part, by several molecules such as Sonic hedgehog and Wnts. Recently, soluble Wnt antagonists have been identified. Among those examined only Frzb1 was found to be expressed in the presomitic mesoderm and newly formed somites and thus its possible role in regulating myogenesis was investigated in detail. When presomitic mesoderm or newly formed somites were cultured with axial structures and surface ectoderm on a feeder layer of C3H10T1/2 cells expressing Frzb1, myogenesis was abolished or severely reduced in presomitic mesoderm and the three most recently formed somites. In contrast, no effect was observed on more mature somites. Inhibition of myogenesis did not appear to be associated with increased cell death since the final number of cells in the explants grown in the presence of Frzb1 was only slightly reduced in comparison with controls. In order to examine the possible function of Frzb1 in vivo, we developed a method based on the overexpression of the soluble antagonist by transient transfection of WOP cells with a Frzb1 expression vector and injection of transfected cells into the placenta of pregnant females before the onset of maternofoetal circulation. Frzb1, secreted by WOP cells, accumulated in the embryo and caused a marked reduction in size of caudal structures. Myogenesis was strongly reduced and, in the most severe cases, abolished. This was not due to a generalized toxic effect since only several genes downstream of the Wnt signaling pathway such as En1, Noggin and Myf5 were downregulated; in contrast, Pax3 and Mox1 expression levels were not affected even in embryos exhibiting the most severe phenotypes. Taken together, these results suggest that Wnt signals may act by regulating both myogenic commitment and expansion of committed cells in the mouse mesoderm.
Borello
U
U
Istituto Pasteur-Cenci Bolognetti, Dipartimento di Istologia ed Embriologia Medica, Università di Roma 'La Sapienza', Via A. Scarpa 14, Italy.
Coletta
M
M
Tajbakhsh
S
S
Leyns
L
L
De Robertis
E M
EM
Buckingham
M
M
Cossu
G
G
D000818:Animals; D001842:Bone and Bones / Q000196:embryology*; D002352:Carrier Proteins; D002454:Cell Differentiation; D004268:DNA-Binding Proteins; D005260:Female; D018507:Gene Expression Regulation, Developmental; D006023:Glycoproteins; D017403:In Situ Hybridization; D047908:Intracellular Signaling Peptides and Proteins; D008648:Mesoderm / Q000378:metabolism*; D051379:Mice; D009124:Muscle Proteins / Q000378:metabolism; D017570:MyoD Protein / Q000378:metabolism; D051787:Myogenic Regulatory Factor 5; D010920:Placenta / Q000378:metabolism*; D011247:Pregnancy; D011506:Proteins / Q000378:metabolism / Q000502:physiology*; D011518:Proto-Oncogene Proteins / Q000378:metabolism*; D019170:Somites / Q000378:metabolism; D015534:Trans-Activators; D014162:Transfection; D051153:Wnt Proteins; D029961:Zebrafish Proteins; D000099176:Noggin Protein; D000099274:Frizzled-Related Proteins
D002352:Carrier Proteins; D004268:DNA-Binding Proteins; D006023:Glycoproteins; D047908:Intracellular Signaling Peptides and Proteins; D009124:Muscle Proteins; D017570:MyoD Protein; D051787:Myogenic Regulatory Factor 5; D011506:Proteins; D011518:Proto-Oncogene Proteins; D015534:Trans-Activators; D051153:Wnt Proteins; D029961:Zebrafish Proteins; D000099176:Noggin Protein; D000099274:Frizzled-Related Proteins; C497418:Myf5 protein, mouse