Expression of Pax-3- and neuroectoderm-inducing activities during differentiation of P19 embryonal carcinoma cells.
A P19 embryonal carcinoma stem cell line carrying an insertion of the E. coli LacZ gene in an endogenous copy of the Pax-3 gene was identified. Expression of the Pax-3/LacZ fusion gene in neuroectodermal and mesodermal lineages following induction of differentiation by chemical treatments (retinoic acid and dimethylsulfoxide) was characterized using this line and is consistent with the previous localization of Pax-3 expression in the embryo to mitotically active cells of the dorsal neuroectoderm and the adjacent segmented dermomyotome. Pax-3/LacZ marked stem cells were also utilized as target cells in mixing experiments with unmarked P19 cells that had been differentiated by pretreatment with chemical inducers. Induction of beta-galactosidase and neuroectodermal markers in the target cells demonstrates that: (1) some differentiated P19 cell derivatives transiently express endogenous Pax-3- and neuroectoderm-inducing activities, (2) undifferentiated target stem cells respond to these activities even in the presence of leukemia inhibitory factor and (3) the endogenous activities can be distinguished from, and are more potent than, retinoic acid treatment in inducing neuroectoderm. These observations demonstrate that P19 embryonal carcinoma cells provide a useful in vitro system for analysis of the cellular interactions responsible for neuroectoderm induction in mammals.
Pruitt
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Roswell Park Cancer Institute, Department of Molecular and Cellular Biology, Buffalo, NY 14263.
D000818:Animals; D015152:Blotting, Northern; D002460:Cell Line; D004268:DNA-Binding Proteins / Q000235:genetics*; D004121:Dimethyl Sulfoxide / Q000494:pharmacology; D004475:Ectoderm / Q000502:physiology*; D004627:Embryonic Induction / Q000187:drug effects / Q000502:physiology*; D005455:Fluorescent Antibody Technique; D015870:Gene Expression / Q000502:physiology*; D008322:Mammals / Q000196:embryology*; D008648:Mesoderm / Q000502:physiology; D009420:Nervous System / Q000196:embryology*; D013724:Teratoma; D014212:Tretinoin / Q000494:pharmacology; D014407:Tumor Cells, Cultured
D004268:DNA-Binding Proteins; D014212:Tretinoin; D004121:Dimethyl Sulfoxide