1995 Fiscal Year Final Research Report Summary
Analysis on the gene expression and the function of FGFs and FGF receptors during induction of differentiation of embryonal carcinoma cells.
| Project/Area Number |
06680701
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
Cell biology
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| Research Institution | Kyoto Sangyo University |
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Principal Investigator |
SEO Misuzu KYOTO SANGYO UNIV.ENGINEERING,ASSOCIATE PROF., 工学部・生物工学科, 助教授 (60211223)
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| Project Period (FY) |
1994 – 1995
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| Keywords | FGF / FGF receptor / neuronal differentiation / Embryonal carcinoma / retinoic acid |
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| Research Abstract |
The changes in the gene expression of fibroblast growth factors (FGFs) and the receptors for FGFs (FGFRs) during retinoic acid-induced neuronal differentiation of mouse embryonal carcinoma -derived P19 cells (EC P19) were examined in this study. EC P19 cells are pluripotent and can be induced to differentiate into neurons and glia cells by exposing cell aggregates to retinoic acid (RA) at 10^<-7> M to 10^<-6>M concentraitons. These neurons resemble morphologically those in the central nervous system, and many neuron-specific genes were shown to be expressed in those cells. Thus, the above system is a good model for studying in vitro what events occur during the critical phase of neuronal differentiation. Gene expression of three FGFs, including FGF2, FGF6, and FGF9 was increased on induction of neuronal differentiation of P19 cells. Among FGFRs, the gene expression of FGFR2 was greatly increased, while that of FGFR1 was constant. Formation of cell aggregates without RA treatment was no
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t enough to induce neuronal differentiation ; most of the cells remained undifferentiated, but a small portion of the cells became heart muscle cells. Under these conditions these FGFs were not induced. In contrast, FGF5 and FGF8 were greatly increased in their expression. Thus, FGF2, FGF6, and FGF9 may play important roles in neuronal development, for example, stimulating growth, survival, and cell-cell adhesion. Among these FGFs, the sequence of mouse FGF9 cDNA expressed in the cells induced by RA was first reported. The murine cDNA showed 92.4% nucleotide sequence homology to the human FGF9 cDNA and 98.2% homology to that of rats. This mouse FGF9 cDNA encoded a polypeptide consisting of 208 amino acids with amino acid sequence identical to that of rats. Only one amino acid was replaced compared to the human homolog. The highly conserved sequence homology of FGF9 suggests its functional importance.
The protein expression of FGF9 during the neuronal differentiation of P19 cells was also examined by staining the cells with anti-FGF9 antibody. Neurons were strongly stained with the antibody, and glial cells were also stained. This study suggests that FGF9 plays important roles in neuronal development, for example, stimulating growth and maintenance of the function of neurons and glial cells. Less
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