Interdepartmental Graduate Program in Neuroscience, Rochester, NY 14642, USA.
J Neurosci Methods. 2010 Jul 15;190(2):171-9. doi: 10.1016/j.jneumeth.2010.05.001. Epub 2010 May 7.
Soluble Abeta 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 Abeta are often unstable and soluble conformer profiles inconsistent. In this study we describe detailed methods for preparing Abeta 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 Abeta-mediated mechanisms underlying AD.
可溶性 Abeta 寡聚体被认为在阿尔茨海默病（AD）的病理生理学中起着关键作用。尽管它们意义重大，但许多研究人员在生成用于体外和体内实验的可靠制剂时遇到了困难。Abeta 的溶液通常不稳定，可溶性构象谱不一致。在这项研究中，我们描述了详细的 Abeta 寡聚体制备方法，这些寡聚体在数周内稳定，并且富含低分子量和高分子量寡聚体形式，包括 56 kDa 形式，该形式与 AD 相关的认知障碍有关。我们使用 Western blot、斑点印迹、原子力显微镜、硫黄素 T 荧光和原代神经元培养毒性测定来表征它们的结构和功能特性。这些合成制剂对于许多研究 AD 中 Abeta 介导的机制的人来说应该是有价值的。
