1991 Fiscal Year Final Research Report Summary
Develpment of the Enzyme Immunoassay System for the Substances Related to the Pathogenesis of Alzheimer's Disease
| Project/Area Number |
02557037
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| Research Category |
Grant-in-Aid for Developmental Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
Neurology
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| Research Institution | Gifu Pharmaceutical University |
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Principal Investigator |
HAYASHI Kyozo Gifu Pharmaceutical University Pharmacy Professor, 薬学部, 教授 (00029935)
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| Co-Investigator(Kenkyū-buntansha) |
HIRATANI Hajime JCR Pharmaceutics Co., Ltd. Biotechnology Research Laboratories, Head, 研究所, 取締役
FURUKAWA Yoshiko Gifu Pharmaceutical University Pharmacy, Associate, 薬学部, 助手 (20219108)
HIRANO Kazuyuki Gifu Pharmaceutical University Pharmacy, Professor, 薬学部, 教授 (90057365)
FURUKAWA Shoei Gifu Pharmaceutical University Pharmacy, Associate Professor, 薬学部, 助教授 (90159129)
NISHITANI Hiroshi Utano National Hospital, President, 宇多野病院, 院長
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| Project Period (FY) |
1990 – 1991
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| Keywords | Alzheimer's disease / cholinergic nerve cells / degeneration / diagnosis / enzyme immunoassay (EIA) / human fibroblast (WS-1) cells / nerve growth factor (NGF) / anti-recombinant human NGF antibody IgG |
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| Research Abstract |
Nerve growth factor(NGF)receptor MRNA has been found to be widely distributed throughout the human central nervous system with the highest levels in the basal f orbrain. This finding suggests that NGF may have function as a retrograde trophic messenger for basal forbrain magnocellular cholinergic nerve cells. The degeneration of the latter constitutes one of the main features of Alzheimer's disease and may be responsible for some of the impairment that characterized this disease. However, little is known about the relationship between Alzheimer's disease and the NGF and other neurotrophic factor levels. This main reason seems to be lack of abundance of these factors in most tissues, and lack of a method to reliably detect the protein levels of these factors including NGF.
Purpose of this research was thus to develop the highly sensitive, specific enzyme immunoassay(EIA)systems for various neurotrophic factors related to the pathogenesis of Alzheimer's disease and to apply these develope
… More
d EIA systems for the diagnosis of the Alzheimer's disease.
Mfajor results are summarized as follows ;
(1)we prepared anti-recombinant human NGF antibody IgG and characterized its property immunologically. This antibody IgG reacted with some animal NGFS, especially with bovine NGF, on immunodiffusion analysis.
(2)Using this antibody IgG, we developed a sensitive two-site EIA system for human NGF, based on the biotin-streptoavidin system. NGF at a concentration as low as 0.02 pg/well(corresponding to 8xlO-19 mol)could be measured with high reproducibility. With this EIA, the detection limit of other, mammalian NGFs was reduced in parallel with the degree of decrease in amino acid seauence homology between them and human NGF.
(3)The NGF in human plasma was found to be 5-10 pg/ml when a 100 mul aliquot of the sample was used.
(4)The specificity of this system was examined using human neurotrophic factor(NT)-3. NT-3 did not show the crossreactivity in this system until the concentration of 10 ng/MI.
(5)Human fibroblast(14S-l)cells were found to synthesize and secrete human NGF in culture and the synthesis/secretion-was regulated in a growth phase-dependent manner.
Preparation of a kit for the measurement of human NGF level in the sera of the patients with Alzheimer's disease. Less
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[Publications] akashi Chijiwa,Atsushi Mishima,Masatoshi Hasegawa,Maroru Sano,Kyozo Hayashi,Tsutomu Inoue,Kenji Naito,Tadashi Toshioka,Hiroyoshi Hidaka: "Inhibition of Forskolin-Induced Neurite Outgrowth and Protein phosphorylation by a Newly Synthesized Selective Inhibitor of Cyclic AMP-Dependent Protein Kinase,N〔2-(p-bromocinnamylamino)-etyl〕-5-isoquinelinesulfinamide (H-89),of PC12D pheochromocytoma." Journal of Biological Chemistry. 265. 5267-5272 (1990)
Description
「研究成果報告書概要(和文)」より
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[Publications] Takashi Chijiwa, Atsushi Mishima, Masatoshi Hasegawa, Mamoru Sano, Kyozo Hayashi, Tsutomu Inoue, Kenji Naito, Tadashi Toshioka, and Hiroyoshi Hidaka: "Inhibition of forskolin-induced neurite outgrowth and protein phosphorylation by a newly synthesized selective inhibitor of cyclic AMP-dependent protein kinase, N-[2-(p-bromocinnamylamino) ethyl]-5-isoquinolinesulfinamide (H-89), of PC12D pheochromocytoma." J. Biol. Chem.265(9). 5267-5272 (1990)
Description
「研究成果報告書概要(欧文)」より
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