An optimized method for enrichment of whole brain-derived extracellular vesicles reveals insight into neurodegenerative processes in a mouse model of Alzheimer's disease.
An optimized method for enrichment of whole brain-derived extracellular vesicles reveals insight into neurodegenerative processes in a mouse model of Alzheimer's disease.
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DOI：
10.1016/j.jneumeth.2018.05.022
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发表时间：
2018-09-01
影响因子：
3
通讯作者：
Meckes DG Jr
中科院分区：
文献类型：
--
作者：
Hurwitz SN;Sun L;Cole KY;Ford CR 3rd;Olcese JM;Meckes DG Jr
阿尔茨海默病（AD）是老年人痴呆的主要原因，并且在过去几十年中患病率急剧增加。然而，许多问题仍然围绕着AD的病因。最近，细胞外囊泡（EV）的运输蛋白质，脂质和核酸从细胞到细胞已涉及在错误折叠的蛋白质的清除和繁殖。研究EV在AD进展中的作用及其潜在的诊断价值可能有助于理解和治疗AD。然而，分离脑源性EV的挑战在一定程度上阻碍了这些研究。在这里，我们提供了一种优化的方法，用于纯化脑源性EV的碘克沙醇浮选密度梯度质谱分析。我们证明了这些囊泡的纯度和EV蛋白的富集相比，沉降梯度分离的囊泡。此外，来自健康和AD小鼠大脑的脑源性EV的比较蛋白质组学分析揭示了囊泡含量的差异，包括参与衰老，免疫应答和氧化还原维持的蛋白质。这些变化为AD相关的神经变性和早期疾病预测的AD潜在生物标志物提供了深入了解。最近的技术已经使用沉降蔗糖梯度从脑组织中分离EV。然而，已经报道了浮选碘克沙醇密度梯度纯化小型EV的优点，并在此进一步证明。总之，这些发现提供了一种严格的技术，用于纯化整个组织来源的EV用于下游分析，并应用这种方法来揭示AD进展和其他神经系统疾病的分子变化。
Alzheimer’s disease (AD) is the major cause of dementia in older adults, and has increased dramatically in prevalence over the past several decades. Yet many questions still surround the etiology of AD. Recently, extracellular vesicles (EVs) that transport protein, lipid, and nucleic acids from cell to cell have been implicated in the clearance and propagation of misfolded proteins. Investigation of EVs in AD progression, and their potential diagnostic utility may contribute to understanding and treating AD. However, the challenges of isolating brain-derived EVs have in part hindered these studies. Here, we provide an optimized method for the purification of brain-derived EVs by iodixanol floatation density gradient for mass spectrometry analysis. We demonstrate the purity of these vesicles and the enrichment of EV proteins compared to sedimentation gradient isolation of vesicles. Moreover, comparative proteomic analysis of brain-derived EVs from healthy and AD mouse brains revealed differences in vesicular content including proteins involved in aging, immune response, and oxidation-reduction maintenance. These changes provide insight into AD-associated neurodegeneration and potential biomarkers of AD for early disease prediction. Recent techniques have used sedimentation sucrose gradients to isolate EVs from brain tissue. However, the advantages of floatation iodixanol density gradient purification of small EVs have been reported and are further demonstrated here. Together these findings offer a rigorous technique for purifying whole tissue-derived EVs for downstream analyses, and application of this approach to uncovering molecular changes in AD progression and other neurological conditions.
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DOI：
10.1083/jcb.200207113
发表时间：
2003-01-06
期刊：
The Journal of cell biology
影响因子：
--
作者：
Ehehalt R;Keller P;Haass C;Thiele C;Simons K
通讯作者：
Simons K
影响因子：
4.2
作者：
Dinkins MB;Dasgupta S;Wang G;Zhu G;Bieberich E
通讯作者：
Bieberich E
影响因子：
82.9
作者：
Alonso, AD;GrundkeIqbal, I;Iqbal, K
通讯作者：
Iqbal, K
影响因子：
4.8
作者：
Jiang, RF;Gao, BC;Eisenberg, E
通讯作者：
Eisenberg, E
影响因子：
5.4
作者：
Hurwitz, Stephanie N.;Nkosi, Dingani;Meckes, David G., Jr.
通讯作者：
Meckes, David G., Jr.