| Project/Area Number |
18500264
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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 |
Nerve anatomy/Neuropathology
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| Research Institution | Kobe University |
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Principal Investigator |
KIKKAWA Satoshi Kobe University, Grad. School of Med, Assistant professor (90244681)
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| Co-Investigator(Kenkyū-buntansha) |
TERASHIMA Toshio Kobe Univ, Grad. School of Med, Professor (20101892)
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| Project Period (FY) |
2006 – 2007
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| Project Status |
Completed (Fiscal Year 2007)
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| Budget Amount *help |
¥4,080,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥480,000)
Fiscal Year 2007: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
Fiscal Year 2006: ¥2,000,000 (Direct Cost: ¥2,000,000)
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| Keywords | Fukuvama-type muscular dystrophy / fukutin / cetral nervous system development / 中枢神経発生障害 / 動物モデル |
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| Research Abstract |
We cloned and sequenced the complete coding region of the zebrafish fukutin gene. Zebrafish fukutin showed a significant degree of sequence homology to mouse and human fukutin. By whole mount in situ hybridization, fukutin mRNA was detected in a wide variety of tissues, including the brain and the spinal cord, as early as 4 hours post-fertilization. We also examined expression patterns of zebrafish orthologs of other muscular dystrophy genes causing central nervous system defects, namely LARGE and POMGnT1. They were also expressed in a variety of tissues from the early developmental stage, implying that sugar-modifying enzymes play important roles in zebrafish CNS development as well.
Next, we analyzed effects of knocking-down Fukutin protein expression on embryogenesis by morpholino antisense oligonucleotide (MO) microinjection into fertilized zebrafish eggs. Injected embryos (morphants) showed a broad range of developmental defects including skeletal and cardiac muscle defects in a dose-dependent manner, reflecting the broad expression of fukutin. We then looked into CNS defects in fukutin morphants using transgenic zebrafish expressing a fluorescent protein either in motor neurons (isl-GFP) or the entire differentiated neuronal pool (HuC-Kaede). Contrary to our expectations, we have not yet been able to find apparent defects of CNS development in both transgenic embryos injected with fukutin MO in spite of gross developmental defects. Further study in more detail is being undertaken for histological analysis.
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