| Co-Investigator(Kenkyū-buntansha) |
KATSUMATA Yoshinao Nagoya University, Graduate School of Medicine, Professor, 大学院・医学系研究科, 教授 (30109326)
YAMAMOTO Toshimichi Nagoya University, Graduate School of Medicine, Associate Professor, 大学院・医学系研究科, 助教授 (50260592)
TAMAKI Keiji Kyoto University, Graduate School of Medicine, Professor, 大学院・医学研究科, 教授 (90217175)
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| Budget Amount *help |
¥3,400,000 (Direct Cost: ¥3,400,000)
Fiscal Year 2005: ¥1,300,000 (Direct Cost: ¥1,300,000)
Fiscal Year 2004: ¥2,100,000 (Direct Cost: ¥2,100,000)
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| Research Abstract |
We selected eight genes related to collagen metabolism such as prolyl 4-hydroxylase (PH), lysyl hydroxylase (LH), procollagen N-proteinase (PNP), procollagen C-proteinase (PCP), prolyl 4-hydroxylase, alpha polypeptide, type 2 (P4HA2), lysyl oxidase (LOX), collagen, type I, alpha-1 (COL1A1) and matrix metalloproteinase 1 (collagenase 1; MMP1) in the first fiscal year, and seven genes of which changes are speculated in skin tissues such as elastin (ELN) constituent to elastic fibers, β-glucocerebrosidase (GBA), sphingomyelinase (SMPD1) and acid ceramidase 1 (ASAH1) related to ceramid metabolism, and tyrosinase (TYR), 5,6-dihydroxy-indole-2-carboxylic acid oxidase (TYRP1), and dopachrome tautomerase (DCT) related to melanin biosynthesis in the second year. A few kinds of PCR primer sets and pyrosequencing primers for the promoter and/or enhancer regions at each gene were designed, PCR reactions were performed using those primer sets with bisulfited template DNA extracted from abdomimal skin tissues collected at autopsy from 4 male and female individuals whose ages ranged from 0 to 80, and then the pyrosequencing reactions were performed. Consequently, only at the second CpG site in TYR-F1 region when using the S2 primer, a strong negative direct proportion between age and methylation were shown. In other almost all regions, no changes of methylation among all samples, or even if some significant changes between samples were observed, those were not concerned to the ages. In the present study, although we could not finally find the clear relationship between methylation at CpG sites and age with skin tissues, it was suggested that some methylated CpG sites related to age might exist because some different degrees of methylation were obsereved in normal tissues.
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