title: Aggregated P19 mouse embryonal carcinoma cells as a simple in vitro model to study the molecular regulations of mesoderm formation and axial elongation morphogenesis.
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.
authors.affiliation: Department of Anatomy, Biochemistry and Physiology, Institute for Biogenesis Research, University of Hawaii School of Medicine, Honolulu, Hawaii 96813, USA. marikawa@hawaii.edu
authors.forename: Yusuke
authors.initials: Y
authors.lastname: Marikawa
authors.forename: Dana Ann A
authors.initials: DA
authors.lastname: Tamashiro
authors.forename: Toko C
authors.initials: TC
authors.lastname: Fujita
authors.forename: Vernadeth B
authors.initials: VB
authors.lastname: Alarcón
mesh_terms: D000818:Animals; D001483:Base Sequence; D019521:Body Patterning; D018236:Carcinoma, Embryonal / Q000235:genetics / Q000378:metabolism / Q000473:pathology*; D002449:Cell Aggregation; D002454:Cell Differentiation; D045744:Cell Line, Tumor; D017931:DNA Primers / Q000235:genetics; D047108:Embryonic Development / Q000235:genetics / Q000502:physiology; D005260:Female; D018507:Gene Expression Regulation, Developmental; D008648:Mesoderm / Q000166:cytology / Q000196:embryology* / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D008811:Mice, Inbred DBA; D008954:Models, Biological; D009024:Morphogenesis / Q000235:genetics / Q000502:physiology; D011247:Pregnancy; D034622:RNA Interference; D015398:Signal Transduction; D051153:Wnt Proteins / Q000037:antagonists & inhibitors / Q000235:genetics; D000071818:Wnt-5a Protein; D060508:Wnt3 Protein; D060509:Wnt3A Protein; D051176:beta Catenin / Q000037:antagonists & inhibitors / Q000235:genetics; D054460:rho-Associated Kinases / Q000378:metabolism
chemical_list: C495267:CTNNB1 protein, mouse; D017931:DNA Primers; D051153:Wnt Proteins; D000071818:Wnt-5a Protein; D060508:Wnt3 Protein; C555508:Wnt3 protein, mouse; D060509:Wnt3A Protein; C555511:Wnt3a protein, mouse; C438615:Wnt5a protein, mouse; D051176:beta Catenin; D054460:rho-Associated Kinases