2000 Fiscal Year Annual Research Report

自己免疫疾患ランバート・イートン筋無力症モデル動物の開発と病因メカニズムの解析

Research Project

Project/Area Number	12557216
Research Institution	The University of Tokyo
Principal Investigator	桐野豊東京大学, 大学院・薬学系研究科, 教授 (10012668)
Co-Investigator(Kenkyū-buntansha)	佐藤一紀福岡女子大学, 人間環境学部, 教授 (10326473) 渡辺恵東京大学, 大学院・薬学系研究科, 助手 (80302610) 川原茂敬東京大学, 大学院・薬学系研究科, 助教授 (10204752) 川上善之エーザイ(株), 筑波探索研究所, 主幹研究員
Keywords	ランバート・イートン筋無力症 / 神経筋接合部 / 自己免疫疾患 / カルシウムチャネル / ダウンレギュレーション / internalization / モデル動物
Research Abstract	1.P型Caチャネルがランバート・イートン筋無力症(LEMS)患者の自己抗体によりダウンレギュレーションされる機構として,チャネルのinternalizationによるという仮説を立て,これを検証する試みを行った.P型Caチャネルに特異的な毒素ω-agatoxin IVAをFluorescein 系(FITC),およびTetramethy lrhodamine(TMRD)でラベルしたプローブを作成した.これらをP型Caチャネルを発現している小細胞型肺ガン細胞(SCLC)に適用したところ,FITC,TMRDラベルのプローブは細胞の自家蛍光と波長帯が重なり,微弱なCaチャネルに結合したプローブの観測には不適当であった.Cy-5によるラベルでは,自家蛍光の影響は回避できたが,蛍光強度が著しく減少したことから,ラベル化反応において何らかの副反応が生じているものと考えられた.そこで,確実に1個のラベルが入ったプローブを作成するために,毒素ペプチドのN末端にCysを導入後,NEMタイプのCy-5を反応させることを考え,現在合成中である. 2.上記1の実験に必要なツールとして,ポリクローナル抗P型Caチャネルペプチド抗体を作成するために,P型Caチャネルの予測トポロジーに基づいて,細胞外ドメインのペプチドで抗原性の高いセグメントを合成し,ウサギに免疫した.これは,また,LEMSモデル動物の作成法の一つでもある.

Research Products
(12 results)

All Other

All Publications (12 results)

[Publications] Inoue,T. et al.: "Phase-dependent filtering of sensory information in the oscillating olfactory center of a terrestrial mollusk"J.Neurophysiol.. 84. 1112-1115 (2000)
[Publications] Tomita,S. et al.: "PDZ domain-dependent suppression of NF-κB/p65-inducedA_42 production by a neuron specific XI I-like protein"J.Biol.Chem.. 275. 13056-13060 (2000)
[Publications] Iijima,K. et al.: "Neuron-specific phosphorylation of Alzheimer's_-amyloid precursor protein by Cdk5"J.Neurochem. 75. 1085-1091 (2000)
[Publications] Lee,D. et al.: "Regulation of XI IL-dependent AP P metabolism by XB51, a novel XI IL-binding protein"J.Biol.Chem.. 275. 23134-23138 (2000)
[Publications] Lee,D. et al.: "Rapid transbilaver phospholipid redistribution associated with exocvtotic release of neurotransmitters from cholinergic nerve terminals isolated from electric ray Narke japonica"Neurosci.Lett.. 291. 21-24 (2000)
[Publications] Toda,S. et al.: "Image analysis of olfactory responses in the procerebrum of the terrestrial slug Limax marginatus."J.Exp.Biol.. 203. 2895-2905 (2000)
[Publications] Nakaya,T. et al.: "Identification and expression of a novel gene in odor-taste associative learning in the terrestrial slug"Genes Cells. 6. 43-56 (2001)
[Publications] Watanabe,S. et al.: "Modulation of oscillatory neural activities by cholinergic activation of interneurons in the olfactory center of a terrestrial slug"Brain Res.. (in press). (2001)
[Publications] Kishimoto,Y. et al.: "Classical eyeblink conditioning in glutamate receptor subunit d2 mutant mice is impaired in delay paradigm but not in trace paradigm"Eur.J.Neurosci.. (in press). (2001)
[Publications] Kishimoto,Y. et al.: "Long-trace interval eyeblink conditioning is impaired in mutant mice lacking NMDA receptor subunit 1"Eur.J.Neurosci.. (in press). (2001)
[Publications] Shindo,K. et al.: "Solution structure of micelle-bound H5 peptide (427-452): A primary structure corresponding to the pore-forming region of the voltage dependent potassium channel"Biochim.Biophys.Acta. (in press). (2001)
[Publications] Satoh,Y. et al.: "β-Amyloid peptides inhibit acetylcholine release from cholinergic presynaptic nerve endings isolated from an electric ray"Neurosci.Lett. (in press). (2001)

2000 Fiscal Year Annual Research Report

自己免疫疾患ランバート・イートン筋無力症モデル動物の開発と病因メカニズムの解析

Principal Investigator

桐野 豊 東京大学, 大学院・薬学系研究科, 教授 (10012668)

Research Products

[Publications] Inoue,T. et al.: "Phase-dependent filtering of sensory information in the oscillating olfactory center of a terrestrial mollusk"J.Neurophysiol.. 84. 1112-1115 (2000)

[Publications] Tomita,S. et al.: "PDZ domain-dependent suppression of NF-κB/p65-inducedA_42 production by a neuron specific XI I-like protein"J.Biol.Chem.. 275. 13056-13060 (2000)

[Publications] Iijima,K. et al.: "Neuron-specific phosphorylation of Alzheimer's_-amyloid precursor protein by Cdk5"J.Neurochem. 75. 1085-1091 (2000)

[Publications] Lee,D. et al.: "Regulation of XI IL-dependent AP P metabolism by XB51, a novel XI IL-binding protein"J.Biol.Chem.. 275. 23134-23138 (2000)

[Publications] Lee,D. et al.: "Rapid transbilaver phospholipid redistribution associated with exocvtotic release of neurotransmitters from cholinergic nerve terminals isolated from electric ray Narke japonica"Neurosci.Lett.. 291. 21-24 (2000)

[Publications] Toda,S. et al.: "Image analysis of olfactory responses in the procerebrum of the terrestrial slug Limax marginatus."J.Exp.Biol.. 203. 2895-2905 (2000)

[Publications] Nakaya,T. et al.: "Identification and expression of a novel gene in odor-taste associative learning in the terrestrial slug"Genes Cells. 6. 43-56 (2001)

[Publications] Watanabe,S. et al.: "Modulation of oscillatory neural activities by cholinergic activation of interneurons in the olfactory center of a terrestrial slug"Brain Res.. (in press). (2001)

[Publications] Kishimoto,Y. et al.: "Classical eyeblink conditioning in glutamate receptor subunit d2 mutant mice is impaired in delay paradigm but not in trace paradigm"Eur.J.Neurosci.. (in press). (2001)

[Publications] Kishimoto,Y. et al.: "Long-trace interval eyeblink conditioning is impaired in mutant mice lacking NMDA receptor subunit 1"Eur.J.Neurosci.. (in press). (2001)

[Publications] Shindo,K. et al.: "Solution structure of micelle-bound H5 peptide (427-452): A primary structure corresponding to the pore-forming region of the voltage dependent potassium channel"Biochim.Biophys.Acta. (in press). (2001)

[Publications] Satoh,Y. et al.: "β-Amyloid peptides inhibit acetylcholine release from cholinergic presynaptic nerve endings isolated from an electric ray"Neurosci.Lett. (in press). (2001)

桐野豊東京大学, 大学院・薬学系研究科, 教授 (10012668)