| Project/Area Number |
63570029
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
神経解剖学
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| Research Institution | SHIGA UNIVERSITY OF MEDICAL SCIENCE |
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Principal Investigator |
KIMURA Hiroshi SHIGA UNIV. OF MEDICAL SCIENCE, DIV. OF NEUROANATOMY, PROFESSOR, 分子神経生物学研究センター, 教授 (40079736)
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| Co-Investigator(Kenkyū-buntansha) |
TOOYAMA Ikuo SHIGA UNIV. OF MEDICAL SCIENCE, DIV. OF NEUROANATOMY, ASSISTANT, 分子神経生物学研究センター, 助手 (20207533)
樋口 彰彦 滋賀医大, 医学部, 助手 (80198986)
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| Project Period (FY) |
1988 – 1989
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| Project Status |
Completed (Fiscal Year 1989)
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| Budget Amount *help |
¥2,200,000 (Direct Cost: ¥2,200,000)
Fiscal Year 1989: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 1988: ¥1,400,000 (Direct Cost: ¥1,400,000)
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| Keywords | Antibody to acetylcholine / Immunohistochemistry / Brain / Cholinergic system / Antigen preparation / 免疫組織学 / コリン神経 |
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| Research Abstract |
Central cholinergic systems have been extensively studied by immunohistochemistry for choline acetyltransferase. a synthesizing enzyme of acetylcholine. Although the method, initiated by a group including the head investigator, provides neuroanatomical knowledge of the cholinergic system in the brain, it is not possible to study functional dynamic state of cholinergic neurons. For this purpose, antibodies directly against neurotransmitters are promising, as seen in the case of serotonin and GABA. In this respect, the head investigators aimed to produce antibody to acetylcholine. So far reported, available acetylcholine antiserum cross-reacts with various choline derivatives. Our strategy to product anti-acetylcholine IgG is to establish a reliable antigen. Since acetylcholine molecule itself is too small to stimulate the immune system of animals receiving immunization, we designed a haptenic antigen for acetylcholine. The haptenic antigen we found to be valuable can be synthesized by reacting succynylcholine chloride with bovine serum albumin via a bridge of glutaraldehyde coupling. The best antiserum, reacted strongly with acetylcholine, did not show any detectable cross-reaction with other choline derivatives known to exist in the body. Even with the antibody, no immunohistochemical staining is obtained in rat brain sections prepared with a commonly used fixation procedure. Further investigation is required to establish a tissue fixation method which is suitable to detect endogenous acetylcholine by using the antibody produced in this study.
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