An improved method for generating consistent soluble amyloid-beta oligomer preparations for in vitro neurotoxicity studies.
An improved method for generating consistent soluble amyloid-beta oligomer preparations for in vitro neurotoxicity studies.
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DOI：
10.1016/j.jneumeth.2010.05.001
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发表时间：
2010-07-15
影响因子：
3
通讯作者：
Bowers, William J.
中科院分区：
文献类型：
--
作者：
Ryan, Deborah A.;Narrow, Wade C.;Federoff, Howard J.;Bowers, William J.
可溶性Aβ低聚物被认为在阿尔茨海默病（AD）病理生理学中发挥关键作用。尽管它们的重要性，许多研究人员遇到困难，在体外和体内实验产生可靠的准备。Aβ的溶液通常是不稳定的，并且可溶性构象谱不一致。在这项研究中，我们描述了制备Aβ寡聚体的详细方法，这些寡聚体可稳定数周，并富含低分子量和高分子量寡聚体形式，包括56-kDa形式，这是一种与AD相关的认知障碍有关的构象异构体。我们使用蛋白质印迹，斑点印迹，原子力显微镜，硫磺素T荧光，和原代神经元培养毒性试验表征其结构和功能特性。这些合成制剂应该证明对许多研究Aβ介导的AD机制有价值。
Soluble Aβ oligomers are recognized as playing a key role in Alzheimer’s disease (AD) pathophysiology. Despite their significance, many investigators encounter difficulty generating reliable preparations for in vitro and in vivo experiments. Solutions of Aβ are often unstable and soluble conformer profiles inconsistent. In this study we describe detailed methods for preparing Aβ oligomers that are stable for several weeks and are enriched for low and high molecular weight oligomeric forms, including the 56-kDa form, a conformer implicated in AD-related cognitive impairment. We characterize their structural and functional properties using Western blot, dot blot, atomic force microscopy, Thioflavine T fluorescence, and primary neuronal culture toxicity assays. These synthetic preparations should prove valuable to many studying Aβ-mediated mechanisms underlying AD.
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影响因子：
15.1
作者：
Kayed R;Head E;Sarsoza F;Saing T;Cotman CW;Necula M;Margol L;Wu J;Breydo L;Thompson JL;Rasool S;Gurlo T;Butler P;Glabe CG
通讯作者：
Glabe CG
影响因子：
25
作者：
Cleary, JP;Walsh, DM;Ashe, KH
通讯作者：
Ashe, KH
影响因子：
16.2
作者：
Oddo, S;Billings, L;LaFerla, FM
通讯作者：
LaFerla, FM
影响因子：
4.7
作者：
Lambert, MP;Viola, KL;Klein, WL
通讯作者：
Klein, WL
DOI：
10.1073/pnas.95.11.6448
发表时间：
1998-05-26
影响因子：
11.1
作者：
Lambert, MP;Barlow, AK;Klein, WL
通讯作者：
Klein, WL