| Budget Amount *help |
¥3,900,000 (Direct Cost: ¥3,900,000)
Fiscal Year 2002: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 2001: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 2000: ¥2,500,000 (Direct Cost: ¥2,500,000)
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| Research Abstract |
Phenotype of genetic polydactly/archinencephaly mouse (Pdn/Pdn) is similar to Greig cephalopolysyndactyly syndrome (GCPS), whose responsible gene is GLI3. Suppression of Gli3 gene expression had been observed in Pdn/Pdn. So, the responsible gene of Pdn/Pdn has been considered to be Gli3, but mutation point has not been demarcated.
In this research project, it was determined that 5442 of a transposon was inserted in the position 41286/41287 of intron 3 of Gli3 gene in Pdn mouse. This transposon has 317-bp long terminal repeats in both ends. Forward and reverse PCR primers were constructed in the intron 3 near the insertion point, and forward primer in the transposon was also a constructed. Then we could discriminate +/+, Pdn/+, Pdn/Pdn embryos with the PCR product by these primers. Using this rapid PCR method, we can determine the genotypes of the young embryos.
Gli3 gene expression in Pdn/+ was suppressed to about 60% for +/+, and that in Pdn/Pdn was 20-30% for +/+ through all the embryonic and neonatal periods examined, but stage dependency of gene expression was not observed. Shh has been considered to be antagonist of Gli3. So, we speculated that Shh would be overexpressed in the Pdn/Pdn mouse embryos whose Gli3 gene expression was suppressed. Shh expression was different among genotypes only by on day of 9 of gestation.
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