人阿尔茨海默病脑中的可溶性淀粉样β聚集物。
Soluble Amyloid-beta Aggregates from Human Alzheimer's Disease Brains.
机构信息
Department of Neurology, 660 South Euclid Avenue, Box 8111, Washington University, St. Louis, Missouri, USA.
Department of Biomedical Engineering, Washington University, St. Louis, Missouri, USA.
出版信息
Sci Rep. 2016 Dec 5;6:38187. doi: 10.1038/srep38187.
Soluble amyloid-beta (Aβ) aggregates likely contribute substantially to the dementia that characterizes Alzheimer's disease. However, despite intensive study of in vitro preparations and animal models, little is known about the characteristics of soluble Aβ aggregates in the human Alzheimer's disease brain. Here we present a new method for extracting soluble Aβ aggregates from human brains, separating them from insoluble aggregates and Aβ monomers using differential ultracentrifugation, and purifying them >6000 fold by dual antibody immunoprecipitation. The method resulted in <40% loss of starting material, no detectible ex vivo aggregation of monomeric Aβ, and no apparent ex vivo alterations in soluble aggregate sizes. By immunoelectron microscopy, soluble Aβ aggregates typically appear as clusters of 10-20 nanometer diameter ovoid structures with 2-3 amino-terminal Aβ antibody binding sites, distinct from previously characterized structures. This approach may facilitate investigation into the characteristics of native soluble Aβ aggregates, and deepen our understanding of Alzheimer's dementia.
可溶性淀粉样蛋白-β (Aβ) 聚集体可能是导致阿尔茨海默病痴呆的主要原因。然而，尽管对体外制剂和动物模型进行了深入研究，但对于人类阿尔茨海默病大脑中可溶性 Aβ 聚集体的特征仍知之甚少。在这里，我们提出了一种从人脑提取可溶性 Aβ 聚集体的新方法，使用差速离心法将其与不溶性聚集体和 Aβ 单体分离，并通过双抗体免疫沉淀法进行>6000 倍的纯化。该方法的起始材料损失<40%，单体 Aβ 无明显的体外聚集，可溶性聚集体大小无明显的体外改变。通过免疫电子显微镜观察，可溶性 Aβ 聚集体通常呈现为 10-20 纳米直径的椭圆形结构簇，具有 2-3 个氨基末端 Aβ 抗体结合位点，与以前表征的结构不同。这种方法可能有助于研究天然可溶性 Aβ 聚集体的特征，并加深我们对阿尔茨海默病痴呆的理解。
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