Large aggregates are the major soluble Aβ species in AD brain fractionated with density gradient ultracentrifugation.
Large aggregates are the major soluble Aβ species in AD brain fractionated with density gradient ultracentrifugation.
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DOI：
10.1371/journal.pone.0032014
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发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
Pettersson FE
中科院分区：
文献类型：
--
作者：
Sehlin D;Englund H;Simu B;Karlsson M;Ingelsson M;Nikolajeff F;Lannfelt L;Pettersson FE
从二聚体到大的原纤维，各种大小的可溶性淀粉样蛋白-β（Aβ）聚集体与阿尔茨海默病（AD）中的神经毒性和突触功能障碍相关。为了研究生物学相关的Aβ种类的性质，在天然条件下，用密度梯度超离心法分离淀粉样β蛋白前体（AβPP）转基因小鼠和AD患者的脑提取物以及合成的Aβ制剂。然后用原子力显微镜（AFM）、ELISA和MTT细胞活力测定分析分级样品。基于AFM外观和对我们的原纤维选择性抗体mAb 158的免疫反应性，将合成Aβ42分为4个级分，级分1中有大的聚集体，级分4中有最小的物质。在MTT试验中，证明来自级分2和3的合成Aβ聚集体毒性最强。在AβPP转基因小鼠脑中，在组分2中发现最丰富的可溶性Aβ物质，主要由Aβ40组成。同样在AD脑中，Aβ主要存在于组分2中，但主要为Aβ42。用mAb 158从免疫学上区分来自组分2的所有生物衍生Aβ与较小物种。因此，发现通过密度梯度超离心天然分离的生物源性可溶性Aβ的主要种类与神经毒性80-500 kDa合成Aβ原纤维的大小相匹配，并且用mAb 158同样检测到。
Soluble amyloid-β (Aβ) aggregates of various sizes, ranging from dimers to large protofibrils, have been associated with neurotoxicity and synaptic dysfunction in Alzheimer's Disease (AD). To investigate the properties of biologically relevant Aβ species, brain extracts from amyloid β protein precursor (AβPP) transgenic mice and AD patients as well as synthetic Aβ preparations were separated by size under native conditions with density gradient ultracentrifugation. The fractionated samples were then analyzed with atomic force microscopy (AFM), ELISA, and MTT cell viability assay. Based on AFM appearance and immunoreactivity to our protofibril selective antibody mAb158, synthetic Aβ42 was divided in four fractions, with large aggregates in fraction 1 and the smallest species in fraction 4. Synthetic Aβ aggregates from fractions 2 and 3 proved to be most toxic in an MTT assay. In AβPP transgenic mouse brain, the most abundant soluble Aβ species were found in fraction 2 and consisted mainly of Aβ40. Also in AD brains, Aβ was mainly found in fraction 2 but primarily as Aβ42. All biologically derived Aβ from fraction 2 was immunologically discriminated from smaller species with mAb158. Thus, the predominant species of biologically derived soluble Aβ, natively separated by density gradient ultracentrifugation, were found to match the size of the neurotoxic, 80–500 kDa synthetic Aβ protofibrils and were equally detected with mAb158.
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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
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作者：
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通讯作者：
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4.2
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Lord, A;Kalimo, H;Nilsson, LNG
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作者：
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通讯作者：
Craievich, AF
影响因子：
120.7
作者：
Näslund, J;Haroutunian, V;Buxbaum, JD
通讯作者：
Buxbaum, JD