The development of the model animals for, and the analysis of the pathogenic mechanism of Lambert-Eaton myasthenic syndrome

Lambert-Eaton肌无力综合征模型动物的建立及发病机制分析

• 批准号: 12557216
• 负责人: KIRINO Yutaka
• 金额: $ 8.38万
• 依托单位: THE UNIVERSITY OF TOKYO
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-12557216/
• 关键词: Calcium channel; Autoimmune disease; Lambert-Eaton myasthenic syndrome; 神経筋接合部; マウス; ダウンレギュレーション; internalization; モデル動物

1. Atropin, a non-selective mAChR antagonist, reduced both mepp amplitude and frequency. By using mAChR-subtype specific knockout mice, it proved that M1 and M3 mAChR are required for transmitter release at NMJ.2. We analyzed whether anti-P/Q-type Ca channel antibody or antisera from LEMS patients reduce the amount of the channel or the Ca influx from the P/Q-type Ca channels expressed stably on the surface of HEK293 cells. Here we successfully generated anti-P/Q-type Ca channel antibody, and showed mat it down-regulated the amount of the channel. It also reduced depolarization-dependent Ca influx.3. It has been thought that LEMS is caused by the reduction in the amount of Ach released from presynaptic terminals in NMJ. To test this, we analyzed the effects of AChE inhibitor E2020 applied in the presence and the absence of K^+ channel blocker 4-aminopyridine. Surprisingly, the effect of the treatment by both of these drugs were not additive, but synergetic against the NMJ transmission.

1. 阿托品是一种非选择性 mAChR 拮抗剂，可降低 mepp 幅度和频率。通过使用mAChR亚型特异性敲除小鼠，证明M1和M3 mAChR是NMJ.2递质释放所必需的。我们分析了LEMS患者的抗P/Q型Ca通道抗体或抗血清是否减少了HEK293细胞表面稳定表达的P/Q型Ca通道的通道量或Ca流入。在这里，我们成功生成了抗P/Q型Ca通道抗体，并显示它下调了通道的数量。它还减少了去极化依赖性钙离子流入。3．人们认为 LEMS 是由 NMJ 突触前末梢释放的 Ach 量减少引起的。为了测试这一点，我们分析了在存在和不存在 K^+ 通道阻滞剂 4-氨基吡啶的情况下应用的 AChE 抑制剂 E2020 的效果。令人惊讶的是，这两种药物的治疗效果不是相加的，而是对 NMJ 传播的协同作用。

项目成果

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