1997 Fiscal Year Final Research Report Summary
Joint Study on the Function of Brain D-Amino Acid Oxidase
| Project/Area Number |
09044315
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| Research Category |
Grant-in-Aid for international Scientific Research
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| Allocation Type | Single-year Grants |
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| Section | Joint Research |
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| Research Field |
General medical chemistry
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| Research Institution | The Institute for Enzyme Research, The University of Tokushima |
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Principal Investigator |
FUKUI Kiyoshi The Inst.for Enzyme Research, The Univ.of Tokushima, Professor, 分子酵素学研究センター, 教授 (00175564)
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| Co-Investigator(Kenkyū-buntansha) |
SAKAI Takashi The Inst.for Enzyme Research, The Univ.of Tokushima, Research Associate, 分子酵素学研究センター, 助手 (80284321)
TOMITA Yumiko The Inst.for Enzyme Research, The Univ.of Tokushima, Research Associate, 分子酵素学研究センター, 助手 (00089913)
YOSEPH Oded Ben Univ.of Tel Aviv, Dept.of Neurobiochemistry, Assistant Professor, 神経生化学部, 講師
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| Project Period (FY) |
1997
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| Keywords | D-amino acid / D-amino acid oxidase / central nervous system / astrocyte |
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| Research Abstract |
D-amino acid oxidase (DAO) is a flavoenzyme with FAD as its prosthetic group that catalyzed the oxidative deamination of a wide range of D-amino acids. In mammals, DAO is found in highest concentrations in kidney, liver and brain. One of the fundamental questions regarding DAO is that of its physiological function, since it woud appear to be an abundant enzyme without a naturally occuringd substrate. Of particular interest are recent reports that D-serine is present in brain and the distribution of DAO in the brain is inversely correlated to that of D-serine.
In search of nervous system specific expression of DAO,RNA blot hybridization analysis of porcine tissues was carried out to show that three mRNA species were expressed in kidney and liver, but only one was detected in brain, indicationg the presence of brain specific regulation of the expression. The reverse transcriptase-coupled PCR analyzes of two human astrocytoma cells derived from cerebral cortex could not detect DAO mRNA.However, double-stranded cDNA of human cerebellar tissue contained high copy number of DAO mRNA.
In ordr to investigate further the expression of cerebral DAO,primary cortical astrocytic cultures were prepared from cerebral cortex isolated from post natal rat pups at day 1-2. The cultures were confluent after 2 weeks in vitro and RNA extraction was performed at day 15. cDNA was synthesized from 1 mug of polyA^+ RNA and PCR analysis was performed. We could detect the amplified DNA of the expected size in cultured cortical astrocytes as well as in cerebellum and kidney. Although DAO has been reporated to be absent in cortex, these results were consistent and reproducible thus indicating the presence of the DAO message in cortical astrocytes.
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