| Project/Area Number |
14570034
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
General anatomy (including Histology/Embryology)
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| Research Institution | RIKEN (The Institute of Physical and Chemical Research) |
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Principal Investigator |
MURAOKA Osamu RIKEN (The Institute of Physical and Chemical Research), Laboratory for vertebrate axis formation, Research scientist, 体軸形成研究チーム, 研究員 (20283765)
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| Project Period (FY) |
2002 – 2003
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| Project Status |
Completed (Fiscal Year 2003)
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| Budget Amount *help |
¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 2003: ¥1,500,000 (Direct Cost: ¥1,500,000)
Fiscal Year 2002: ¥2,000,000 (Direct Cost: ¥2,000,000)
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| Keywords | zebrafish / neural area / kheper / mouse / ES cell / Expression cloning / Zebrafish / ES cell / Mouse / Expression cloning / ゼブラフイツシュ / 神経発生 |
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| Research Abstract |
We screened cDNA libraries using expression screen strategy to identify the factors that regulated the development of the neuroectoderm. We made the plasmid cDNA libraries from zebrafish embryos, mouse embryos, and early differentiated mouse embryonic stem (ES) cells. These were transformed into the E.coli and divided in groups which contained 200 clones each. We prepared the plasmid from each groups and made RNAs for the microinjection. These RNAs were injected into the zebrafish embryos at one to two cell stage. We checked the phenotype by microscopic view and the whole mount in situ hybridization with neural marker probes. As the neural marker, we used six3,eng3,krox20 and kheper. kheper is the new transcription factor which we cloned. kheper is expressed in the whole neuroectoderm and regulated the development of the neural tissue of zebrafish.
We screened 23 x 10^4 clones and found many well known genes. In addition, we succeeded to identify new FGF family gene and new Bcl2 family gene and new wnt family gene. When these are overexpressed in zebrafish embryos, the neuroectoderm were enlarged. The expression of the new fgf was detected in the anterior region of the central nervous system. It is suggested that the new FGF regulates the development of the forebrain.
We are going to do the epistatic analysis of these factors. We would like to know the relationship of these with kheper and how they regulate the development of neural tissue.
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