| Project/Area Number |
11480225
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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 |
Nerve anatomy/Neuropathology
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| Research Institution | Yamagata University |
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Principal Investigator |
GOTO Kaoru Yamagata University, School of Medicine Department of Anatomy and Cell Biology, Professor, 医学部, 教授 (30234975)
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| Co-Investigator(Kenkyū-buntansha) |
NAKANO Tomoyuki Yamagata University, School of Medicine Department of Anatomy and Cell Biology, Assistant professor, 医学部, 助手 (00333948)
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| Project Period (FY) |
1999 – 2001
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| Project Status |
Completed (Fiscal Year 2001)
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| Budget Amount *help |
¥14,000,000 (Direct Cost: ¥14,000,000)
Fiscal Year 2001: ¥2,900,000 (Direct Cost: ¥2,900,000)
Fiscal Year 2000: ¥3,600,000 (Direct Cost: ¥3,600,000)
Fiscal Year 1999: ¥7,500,000 (Direct Cost: ¥7,500,000)
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| Keywords | phospholipid / diacylglycerol / diacylglycerol kinase / neuron / nucleus / signal transduction / second messenger / 脳 / 初代培養神経細胞 / 遺伝子導入 / 核移行シグナル / アンキリンリピート |
|---|
| Research Abstract |
Diacylglycerol kinase (DGK), which catalyzes phosphorylation of lipid second messenger, DG to phosphatidic acid (PA), is thought to be a key enzyme in the regulation of DG levels and, as a result, to be responsible for attenuating the activation of PKC. We have so far cloned several isozymes from rat brain cDNA library, and performed detailed examination of their gene expression in the brain by in situ hybridization histochemistry. In this study we analyzed precise subcellular localization of the isozymes in cDNA-transfected COS cells using green fluorescent protein-fusion expression vector. DGK-alpha was localized diffusely in both cytoplasm and nucleus in the cells. DGK-beta was distributed throughout the cytoplasm in the shape of filamentous structure, which is quite similar to the location of actin filament. DGK-gamma was seen as a reticular structure in the juxtanuclear position, which is coincided with Golgi apparatus marker. DGK-zeta was located mostly in the nucleus, as suggested by the presence of nuclear localizing signal in its primary structure. DGK-epsilon was widespread to the perinuclear area, which resembles the location pattern of endoplasmic reticulum. These data clearly show the heterogeneity of DGK in subcellular localization as well as molecular structure, suggesting that each isozyme may be responsible for the metabolism of DG, which is produced at specific subcellular site upon stimulation. Therefore it is highly conceivable that each DGK isozyme plays a unique role in the signal transduction.
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