低聚β-淀粉样蛋白的制备 1-42和突触可塑性损伤诱导海马脑片
Summary
阿尔茨海默氏病的一个特点是海拔的Aβ<sub> 1-42</sub>肽。在这里，我们提供的协议，为准备合成的Aβ<sub> 1-42</sub>低聚物，损害海马长时程增强，蜂窝关联的内存。此过程是有用的调查Aβ诱导的病理和药物筛选机制。
Abstract
突触连接的减值可能健忘症在阿尔茨海默病（AD）的早期阶段发生的微妙变化的基础。被称为β-淀粉样蛋白（Aβ），在AD中高量生产的一种肽，以减少长时程增强（LTP），蜂窝的学习和记忆相关的。事实上，由Aβ引起的LTP的减值是一个有用的实验范式，研究AD模型和筛选药物能够减轻或恢复这种突触损伤的突触功能障碍。有研究表明，Aβ的产生通过其低聚体形式的LTP中断优先。在这里，我们提供oligomerized合成灌注损害LTP的一个详细的协议到急性海马Aβ1- 42肽。在这段视频中，我们概括为低聚Aβ的准备一步一步程序<sub> 1-42</sub>。然后，我们按照个人的实验中，减少暴露oligomerizedAβ的海马LTP是<sub> 1-42</sub>在控制实验没有Aβ的中，以片相比<sub> 1-42</sub>接触发生了。
Protocol
Discussion
如上所述，在编制低聚Aβ的几个问题可能会导致在受到损害LTP的。评估是否可能发生Aβ的降解或缺乏对Aβ齐聚的方法之一是评估使用的Tris -甘氨酸页/免疫印迹分析和分析超速离心（未在本协议中所述）Aβ的准备。当免疫印迹样品准备non-denaturing/non-reducing条件下，一个成功的准备工作应产生免疫印迹签名具有对应单体及不同的低聚物3的乐队。一个经常发生的问题是在SpeedVac浓缩干燥步?…
Divulgazioni
The authors have nothing to disclose.
Acknowledgements
这项工作是由美国国立卫生研究院国家神经疾病与中风研究所（NINDS）（NS049442）的支持。>
Materials
| Material Name | Tipo | Company | Catalogue Number | Comment |
|---|---|---|---|---|
| Aβ 1-42 (lyophilized) | American Peptide Co. | 62-0-80 | ||
| 1,1,1,3,3,3-Hexafluoro-2-Propanol (HFIP) | Fluka | 52517 | ||
| Dimethylsulfoxide (DMSO) | Biotium | 90082 | ||
| 50mL Polypropylene Centrifuge Tubes | Corning | 05-538-67 | ||
| 1.5ml MaxyClear microtubes Maximum recovery | Axygen | MCT-150-L-C | ||
| Phosphate-Buffered Saline pH 7.4 | Invitrogen-Gibco | 10010-023 | ||
| GASTIGHT Syringes 250 μL | Hamilton | 1725 | ||
| Bath sonicator | Branson | 3510 | ||
| Savant SpeedVac Concentrator System | Various distributors | SC 110A |
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