| Project/Area Number |
02557011
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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 |
General pharmacology
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| Research Institution | Kyoto Prefectural University of Medicine |
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Principal Investigator |
KURIYAMA Kinya Kyoto Pref.Univ.of Med.Pharmacol.,Professor, 医学部, 教授 (20079734)
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| Co-Investigator(Kenkyū-buntansha) |
KATSURA Masashi Kyoto Pref.Univ.of Med.Pharmacol.,Instructor, 医学部, 助手 (80204452)
HIROUCHI Masaaki Kyoto Pref.Univ.of Med.Pharmacol.,Instructor, 医学部, 助手 (70181196)
TAGUCHI Jun-ichi Kyoto Pref.Univ.of Med.Pharmacol.,Instructor, 医学部, 助手 (50188132)
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| Project Period (FY) |
1990 – 1992
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| Project Status |
Completed (Fiscal Year 1992)
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| Budget Amount *help |
¥5,800,000 (Direct Cost: ¥5,800,000)
Fiscal Year 1992: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 1991: ¥1,600,000 (Direct Cost: ¥1,600,000)
Fiscal Year 1990: ¥3,200,000 (Direct Cost: ¥3,200,000)
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| Keywords | GABA_Areceptor antibody / Receptor purification / GABA_B_receptor antibody / Monoclonal antibody / Baclofen affinity column / Immunoaffinity column / Immunohistochemistry / Electron Microscopic immuno-cytochemistry / GABA_A受容体 / GABA_B受容体 / インミュノアフィニティ精製 / 再構成 / 小脳 / GABA_A受容体複合体 / モノクロ-ナル抗体 / mRNA / 純化精製 / cDNAクロ-ニング / ド-パミン作働性ニュ-ロン / 免疫抗体作製 |
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| Research Abstract |
Immunohistochemical procedures for cerebral GABA_A and GABA_B receptors using specific monoclonal antibodies have been developed. The results obtained are as follow. 1)Albin o rabbits were immunized against partially purified GABA_A receptor complex from the bovine brain using baclofen-affinity column, and the specific antibody for GABA_A receptor complex was obtained. 2) Mice were immunized against partially purified GABA_B receptor protein using baclofen-affinity column, and the monoclonal antibody which recognizes 80KDa GABA_B receptor was obtained. 3) The immunoaffinity column using GABA_B receptor monoclonal antibody was obtained, and complete purification of the GABA_B receptor protein(80KDa) was achieved using this immunoaffinity column. 4) The purified GABA_B receptor possessed pharmacological properties as GABA_B receptor and exhibited the inhibition of cAMP formation when it was reconstituted in phospholipid vesicles with purified adenylate cyclase and Gi/Go type of GTP binding protein. 5) Immunohistochemical studies using above GABA_A receptor and GABA_B receptor antibodies revealed that the localization of GABA_A receptor and GABA_B receptors at GABAergic synapses was different. 6) It has been confirmed that these immunohistochemical procedures are useful for the analysis of GABAergic pathways in the brain.
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