Detection of elevated levels of soluble -synuclein oligomers in post-mortem brain extracts from patients with dementia with Lewy bodies
Detection of elevated levels of soluble -synuclein oligomers in post-mortem brain extracts from patients with dementia with Lewy bodies
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DOI：
10.1093/brain/awn349
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发表时间：
2009-04-01
期刊：
影响因子：
14.5
通讯作者：
El-Agnaf, Omar M. A.
中科院分区：
文献类型：
--
作者：
Paleologou, Katerina E.;Kragh, Christine L.;El-Agnaf, Omar M. A.
许多神经退行性疾病包括帕金森病、路易体痴呆（DLB）和多系统萎缩，其特征在于β-突触核蛋白（β-syn）蛋白在受影响的脑区域中的纤维状聚集体的形成和神经元内积聚。这些和其他研究结果表明，-syn在大脑中的积累在这些疾病的发病机制中起着重要作用。然而，最近有报道称，早期淀粉样蛋白聚集体或可溶性低聚物是导致神经变性和神经元细胞死亡的致病物质，而不是后来成熟的原纤维。在这项研究中，我们调查了存在的-syn低聚物在脑裂解液中制备的冷冻死后正常，阿尔茨海默病和DLB患者的大脑。对脑提取物进行高速离心，以去除不可溶的-syn聚集体，然后通过使用FILA-1（一种特异性结合-syn聚集体但不结合-syn单体或tau的抗体）特异性检测上清液中的可溶性低聚物。或-淀粉样蛋白聚集体。使用这种新的酶联免疫吸附测定（ELISA）方法来定量的-syn低聚物的脑提取物中的量，我们的数据清楚地表明，在DLB脑中的可溶性低聚物的-syn的水平增加相比，阿尔茨海默病和控制（P 0.0001）。我们的研究结果提供了强有力的证据，以支持这一论点，即升高的可溶性寡聚体的-syn参与DLB的发病机制。此外，这些发现建立FILA-1作为一个非常敏感的工具，用于检测人脑裂解物中的-syn的寡聚体形式。
A number of neurodegenerative diseases including Parkinsons disease, dementia with Lewy bodies (DLB) and multiple system atrophy are characterized by the formation and intraneuronal accumulation of fibrillar aggregates of -synuclein (-syn) protein in affected brain regions. These and other findings suggest that the accumulation of -syn in the brain plays an important role in the pathogenesis of these diseases. However, more recently it has been reported that early amyloid aggregates or soluble oligomers are the pathogenic species that lead to neurodegeneration and neuronal cell death rather than the later mature fibrils. In this study, we investigated the presence of -syn oligomers in brain lysates prepared from frozen post-mortem brains of normal, Alzheimers disease and DLB patients. The brain extracts were subjected to high speed centrifugation, to remove insoluble -syn aggregates, followed by specific detection of soluble oligomers in the supernatants by employing FILA-1, an antibody that specifically binds to -syn aggregates, but not to -syn monomers, or to tau or -amyloid aggregates. Using this novel enzyme-linked immunosorbent assay (ELISA) method to quantify the amounts of -syn oligomers in the brain extracts, our data clearly show an increase in the levels of soluble oligomers of -syn in the DLB brains compared to those with Alzheimers disease and the controls (P 0.0001). Our findings provide strong evidence to support the contention that elevated soluble oligomers of -syn are involved in the pathogenesis of DLB. Furthermore, these findings establish FILA-1 as a very sensitive tool for the detection of oligomeric forms of -syn in human brain lysates.