| Project/Area Number |
11680755
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Neurochemistry/Neuropharmacology
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| Research Institution | Osaka University |
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Principal Investigator |
NIINOBE Michio Institute for Protein Research, Osaka University, Associate Professor, 蛋白質研究所, 助教授 (80135748)
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| Co-Investigator(Kenkyū-buntansha) |
MOCHIDA Sumiko Tokyo Medical College, Associate Professor, 助教授 (30096341)
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| Project Period (FY) |
1999 – 2000
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| Project Status |
Completed (Fiscal Year 2000)
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| Budget Amount *help |
¥2,600,000 (Direct Cost: ¥2,600,000)
Fiscal Year 2000: ¥1,300,000 (Direct Cost: ¥1,300,000)
Fiscal Year 1999: ¥1,300,000 (Direct Cost: ¥1,300,000)
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| Keywords | Amyloid Precursor Protein / Synaptic Vesicle / Endocytosis / NT-2 Cell / Caspase-3 / Apoptosis / Caenorhabditis elegans / Serine-threonine Kinase / アルツハイマー病 / ラット交感神経節細胞 / エンドサイトーシス / セリン・スレオニンキナーゼ様蛋白質 / カスパーゼ / アデノウィスルベクター |
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| Research Abstract |
We studied on physiological functions of amyloid precursor protein (APP695) in neural cells, and found following results. (1) Postsynaptic responses evoked by a presynaptic action potentials decreased gradually, when the antibody to Cterminal peptide (25 amino acid residues) of APP695 or C-terminal peptide was injected into presynaptic neurons of cholinergic synapses formed between rat sympathetic neurons in culture. The same results were also observed with injection of C-terminal small peptide (8 amino acid residues) relevant to endocytosis. These results suggested that inhibition of the synaptic transmission with the antibody or the C-terminal peptide resulted from inhibition of re-uptake of synaptic vesicles. (2) We analyzed effects of APP695 over-expressed with adenovirus vector into human postmitotic neural cells (NT-2). The results revealed that, in immunohistochemical and biochemical approaches, activated caspase-3 was detected in NT-2 cells at 48 hours after the transfection. Further apoptotic degeneration of NT-2 cells was observed at 72 hours. But the degeneration was blocked by addition of caspase-3 inhibitor into the culture medium. These results suggest that APP695 over-expressed into NT-2 cells induced apoptotic degeneration with activation of caspase-3 by some mechanism. (3) We screened the binding proteins to the extracellular domain in the extract of adult mouse brain using GST-fusion protein or biotinylated peptide as ligands. The results revealed that some proteins with high molecullar weight were found using GST-fusion protein of the extracellular domain (amino acid sequence 1-596) as a ligand, and that mouse homologue of serine-threonine kinase like protein of Caenorhabditis elegans was found using biotinylated peptide (amino acid sequence 66-81) as a ligand.
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