| Project/Area Number |
60870029
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| Research Category |
Grant-in-Aid for Developmental Scientific Research
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| Allocation Type | Single-year Grants |
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| Research Field |
内科学一般
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| Research Institution | HIROSHIMA UNIVERSITY |
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Principal Investigator |
KITO Shozo Hiroshima University School of Medicine, 医学部, 教授 (00010140)
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| Co-Investigator(Kenkyū-buntansha) |
SHIMOYAMA Masanori Hiroshima University School of Medicine, 医学部附属病院, 助手 (60136067)
INAGAKI Shinobu Hiroshima University School of Medicine, 医学部, 講師 (90151571)
YAMAMURA Yasuhiro Hiroshima Uniersity School of Mecidine, 医学部附属病院, 講師 (10106388)
松林 弘明 広島大学, 医学部, 助手 (60165850)
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| Project Period (FY) |
1985 – 1987
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| Project Status |
Completed (Fiscal Year 1987)
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| Budget Amount *help |
¥5,700,000 (Direct Cost: ¥5,700,000)
Fiscal Year 1987: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 1986: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 1985: ¥4,000,000 (Direct Cost: ¥4,000,000)
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| Keywords | Neutotransmitter receptor / invitro Autoradiography / Quantitative analysis / Standard / Neurogenerative disorder / 個体発生 / In vitro autoradiography / 神経伝達物質受容体 / 受容体の絶対数 / 各種変性性神経疾患剖検脳 / 神経伝達物質受容体の絶対数 / オートラジオグラフィー / 標準曲線 / Unnerstallの式 / 画像解析 |
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| Research Abstract |
in vitro Macro-autoradigraphy is the best and only method to determine distrbution of neurotransmitter receptors in each discete region within the central nervous system. The purpose of this research is to establish the method to calculate the absolute number of receptors in autoradiographic images of the brain using tritiated rediactive standards. Actually, Standaedizartion was done by binding of 3H-QNB which is an antabonist of muscatineic acetylcholine receptor. Homogenate of the normal rat brain was prepared adn part of isa was incubated with^3H-QNB to calculate the binding expressed as mol/mg protein. The rest of incubated solution was centriguged and the obtained pellet was applied to a H-sensitive film according to the ordinary process of macro-autoradiography. We utilized the IBAS II computer system of Zeiss company to determine relationship between autoradiographic optical density and radioactivity. As the results, we noticed that there was a linewar relationship between optical density and tissue-bound radioactivity when converted to a In-In scale. With these experiments, we obrained a very good consistence between the absolute number of receptore calculated from in vitro autoradioragraphy and that from brain homogenate binding experiments. This means that the standard prepared by us has a good quality enough to use as a standard of quantitative in vitro autoradiography. Through this rechnique esteblished by us, we examined distributions of various neurotransmitter receptors in the brains of MPTP-induced monkey parkinsonism, human cases of spinocerebellar degenetation and Alzheimer disease, and animal models of these neurological desorders. Besids, ontogenic developments of muscatinic acetylcholine receptor subtypes and neutotensin receptors in rat brains were examined. Throught these experiments, is was confirmed thast quantitative in vitro autoradigrraphy is a reliable and widely applicable experimental technique in neuroscience.
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