1996 Fiscal Year Final Research Report Summary
Screening for the smooth muscle relaxants
| Project/Area Number |
07557310
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| Research Category |
Grant-in-Aid for Scientific Research (A)
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| Allocation Type | Single-year Grants |
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| Section | 試験 |
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| Research Field |
応用薬理学・医療系薬学
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| Research Institution | Gunma University School of Medicine |
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Principal Investigator |
KOHAMA Kazuhiro Gunma University, School of Medicine, Department of Pharmacology, Professor, 医学部, 教授 (30101116)
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| Co-Investigator(Kenkyū-buntansha) |
SAIDA Koichi Bayer Ltd, Chief investigater, 薬理学研究所, 主任研究員
NAKAMURA Akio Gunma University, School of Medicine, Department of Pharmacology, assistant, 医学部, 助手
ISHIKAWA Ryoki Gunma University, School of Medicine, Department of Pharmacology, assistant, 医学部, 助手 (20212863)
OKAGAKI Tsuyoshi Gunma University, School of Medicine, Department of Pharmacology, lecturer, 医学部, 講師 (80185412)
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| Project Period (FY) |
1995 – 1996
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| Keywords | purinergic receptor / capsicin receptor / patch clamp / Cytoskeleton / PC-12 Cell / actin / nerve cell / recombinant protein |
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| Research Abstract |
This research aims to examine the effects of cytoskeletal proteins on the activity of ionotropic receptor. It has been known for many years that Cytochalasin and Colchicine, which destroy cytoskeltons, affected some receptor activities, when they were applied extracellularly. Therefore, we expected that actin and tubilin and their associated proteins may exert regulatory roles to the ionotropic channels.
In 1996, we applied cytochalasin (which destroys actin filaments) and phalloidin (which stabilizes actin filaments) and recorded the changes in the ionic currents of purinergic P_2x_2 by the patch clamp method. Unfortunately we failed to detect any significant effects of the agents.
In 1997, we obtained cDNA coding P_2x_2 receptor by subjecting PC-12 cells to RT-PCR method. We then tried to express the receptor in E.Coli by the pET expression system so that the interaction between P_2x_2 and cytoskeletal proteins was examined. However, E.Coli transfected with the system did not produce P_2x_2.
In 1998. we obtained cDNA coding a capsicin receptor by subjecting dorsal root ganglion to RT-PCR, and inserted the PCR product to pET expression system. E.Coli transfected the system fortunately produced the capsicin receptor. We purified the receptor protein by the column chromatographies in a large amount. Actin-binding activity was detected for the receptor protein by precepitating it together with actin filaments. This preliminary experiment will provide a clue to examine the effect of cytoskeletal proteins on P_2x_2 activity.
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Research Products
(13 results)
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[Publications] Hayashi, K., Ishikawa, R., Ye, L.-H., He, X.-L., Takata, K., Kohama, K.and Shirao, T.: "Modulatory role of drebrin on the cytoskeleton within dendritic spines in the rat cerebral cortex." J.Neurosci.16. 7161-7170 (1996)
Description
「研究成果報告書概要(欧文)」より
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[Publications] Ye, L.-H., Hayakawa, K., Kishi, H., Imamura, M., Nakamura, A., Okagaki, T., Takagi, T., Iwata, A., Tanaka, T.and Kohama, K.: "The structure and function of the actin-binding domain of myosin light chain kinase of smooth muscle." J.Biol.Chem.272. 32182-32189 (1997)
Description
「研究成果報告書概要(欧文)」より
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