| Budget Amount *help |
¥17,810,000 (Direct Cost: ¥13,700,000、Indirect Cost: ¥4,110,000)
Fiscal Year 2014: ¥6,110,000 (Direct Cost: ¥4,700,000、Indirect Cost: ¥1,410,000)
Fiscal Year 2013: ¥5,980,000 (Direct Cost: ¥4,600,000、Indirect Cost: ¥1,380,000)
Fiscal Year 2012: ¥5,720,000 (Direct Cost: ¥4,400,000、Indirect Cost: ¥1,320,000)
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| Outline of Final Research Achievements |
The structure and function of the Pax1 genomic region, which is essential for vertebral column formation, was compared between the mouse and zebrafish by a cross-species genome engineering approach. Transgenic reporter assays revealed that a mouse sclerotome (vertebral anlage)-specific enhancer mPf1 (0.8 kb) did not show enhancer activity in zebrafish embryos, while its zebrafish ortholog zPf1 (0.7 kb) drove stronger sclerotomal expression in mice than the isogenic mPf1. Another sclerotomal enhancer mXe1 (1.7 kb), which is evolutionary conserved in terrestrial vertebrates but not in aquatic vertebrates, drove robust reporter expression in the zebrafish sclerotome. Intriguingly, forced expression of Pax1 driven by the Xe1 enhancer transgene inhibited cartilage formation in zebrafish. The inhibitory role of Pax1 against cartilage formation was further analyzed in detail by forced expression experiments in chicken embryos and primary cultured cells.
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