1998 Fiscal Year Final Research Report Summary
Isolation of Differentially Expressed Genes in Rat Hippocampus after Transient Global Cerebral Ischemia Using PCR mediating Subtractive cDNA Cloning.
| Project/Area Number |
08457361
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Cerebral neurosurgery
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| Research Institution | Hamamatsu University |
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Principal Investigator |
YOKOTA Naoki Hamamatsu Univ.Schl.Med., Assistant of Professor, 医学部附属病院, 助手 (00273186)
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| Co-Investigator(Kenkyū-buntansha) |
UEMURA Kenichi Hamamatsu Univ.Schl.Med., Professor, 医学部, 教授 (60009561)
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| Project Period (FY) |
1996 – 1998
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| Keywords | Hippocampus / Transient Global Cerebral Ischemia / PCR / Subtractive cDNA Cloning / Rat |
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| Research Abstract |
To clarify molecular mechanisms underlying phenomenone in hippocampus after transient cerebral global isehemia, we have isolated differentially expressed genes in rat hippocampus of in vivo transient cerebral global ischemia model using PCR-mediating subtractive cloning. Homology search against database (DNA Data Bank of Japan) revealed that among over 100 fragments which we isolated, up-to 1/2 of these were identical, remains showed homology to known sequences up-loaded and only two displayed no homology. Among identified genes, 10 clones were selected randomly and quantitative RT-PCR analyses were performed. Half of them (5110) were revealed increased expression in 24 hour after ischemia over two times compared to sham operated. These are identified to furin, prosaposin, neutral and basic amino acid transporter (NBAT) synaptotagmin IV and heat shock protein 105, respectively. Furthermore, clonological expression pattern of mRNA of these genes were analyzed and it was revealed that furin had high peak at 6 hr after ischemia but peak of other 4 genes were delayed and had at 24 hr after. It was also suggested that PCR-mediating subtractive cDNA cloning is a very efficient and useful tool for analyzing genes in rat ischemic hippocampus of in vivo transient cerebral global ischemia model.
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