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Isolation of Differentially Expressed Genes in Rat Hippocampus after Transient Global Cerebral Ischemia Using PCR mediating Subtractive cDNA Cloning.

Research Project

Project/Area Number 08457361
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field Cerebral neurosurgery
Research InstitutionHamamatsu University

Principal Investigator

YOKOTA Naoki  Hamamatsu Univ.Schl.Med., Assistant of Professor, 医学部附属病院, 助手 (00273186)

Co-Investigator(Kenkyū-buntansha) UEMURA Kenichi  Hamamatsu Univ.Schl.Med., Professor, 医学部, 教授 (60009561)
Project Period (FY) 1996 – 1998
Project Status Completed (Fiscal Year 1998)
Budget Amount *help
¥7,800,000 (Direct Cost: ¥7,800,000)
Fiscal Year 1998: ¥600,000 (Direct Cost: ¥600,000)
Fiscal Year 1997: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 1996: ¥6,500,000 (Direct Cost: ¥6,500,000)
KeywordsHippocampus / Transient Global Cerebral Ischemia / PCR / Subtractive cDNA Cloning / Rat
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.

Report

(3 results)
  • 1998 Final Research Report Summary
  • 1997 Annual Research Report
  • 1996 Annual Research Report

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Published: 1996-04-01   Modified: 2016-04-21  

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