Mesh : Animals Base Sequence Body Patterning Carcinoma, Embryonal / genetics metabolism pathology Cell Aggregation Cell Differentiation Cell Line, Tumor DNA Primers / genetics Embryonic Development / genetics physiology Female Gene Expression Regulation, Developmental Mesoderm / cytology embryology metabolism Mice Mice, Inbred C57BL Mice, Inbred DBA Models, Biological Morphogenesis / genetics physiology Pregnancy RNA Interference Signal Transduction Wnt Proteins / antagonists & inhibitors genetics Wnt-5a Protein Wnt3 Protein Wnt3A Protein beta Catenin / antagonists & inhibitors genetics rho-Associated Kinases / metabolism

来  源:   DOI:10.1002/dvg.20473   PDF(Sci-hub)   PDF(Pubmed)

Abstract:
Because of their capacity to give rise to various types of cells in vitro, embryonic stem and embryonal carcinoma (EC) cells have been used as convenient models to study the mechanisms of cell differentiation in mammalian embryos. In this study, we explored the mouse P19 EC cell line as an effective tool to investigate the factors that may play essential roles in mesoderm formation and axial elongation morphogenesis. We first demonstrated that aggregated P19 cells not only exhibited gene expression patterns characteristic of mesoderm formation but also displayed elongation morphogenesis with a distinct anterior-posterior body axis as in the embryo. We then showed by RNA interference that these processes were controlled by various regulators of Wnt signaling pathways, namely beta-catenin, Wnt3, Wnt3a, and Wnt5a, in a manner similar to normal embryo development. We further showed by inhibitor treatments that the axial elongation morphogenesis was dependent on the activity of Rho-associated kinase. Because of the convenience of these experimental manipulations, we propose that P19 cells can be used as a simple and efficient screening tool to assess the potential functions of specific molecules in mesoderm formation and axial elongation morphogenesis.
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