Proteomic analysis of the brain tissues from a transgenic mouse model of amyloid β oligomers
Proteomic analysis of the brain tissues from a transgenic mouse model of amyloid β oligomers
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DOI：
10.1016/j.neuint.2012.05.018
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发表时间：
2012-08-01
影响因子：
4.2
通讯作者：
Matsuyama, Shogo
中科院分区：
文献类型：
--
作者：
Takano, Masaoki;Maekura, Kouji;Matsuyama, Shogo
淀粉样蛋白β（A β）寡聚体被认为是阿尔茨海默病（AD）的病因之一。以前，我们在淀粉样蛋白成像中几乎没有显示淀粉样斑块信号的AD患者中确定了淀粉样前体蛋白（APP）的E693 Delta突变。我们产生了表达E693 Delta突变的APP转基因小鼠，发现它们从8月龄开始就具有丰富的A β寡聚体，但即使在24月龄也没有淀粉样斑块，这表明这些小鼠是研究A β寡聚体病理作用的良好模型。为了阐明A β寡聚体是否会影响大脑中的蛋白质组水平，我们检查了12个月大的APP（E693 Delta）转基因小鼠与年龄匹配的非转基因同窝小鼠相比，蛋白质和磷蛋白水平发生了变化。通过双向凝胶电泳（2DE）、SYPRO Ruby和Pro-Q Diamond染色以及随后的质谱技术，我们鉴定了APP（E693 Delta）转基因小鼠海马和大脑皮层中17种蛋白和3种磷蛋白发生显著变化。辅肌动蛋白样蛋白，SH 3结构域结合谷氨酸丰富的样蛋白3和星形胶质细胞磷蛋白PEA-15亚型2的水平降低到低于0.6倍的非转基因同窝仔，而发动蛋白，profilin-2，空泡腺苷三磷酸酶和肌酸激酶B的水平增加到1.5倍以上的非转基因同窝仔。此外，2DE蛋白质印迹法验证了APP（E693 Delta）转基因小鼠中发动蛋白、二氢嘧啶酶相关蛋白2（Dpysl 2）和辅肌动蛋白水平的变化。谷胱甘肽酶和异柠檬酸脱氢酶的水平增加到非转基因同窝出生的1.5倍以上。已鉴定的蛋白质可分为几组，它们参与不同细胞功能的调节，例如细胞骨架及其相互作用蛋白、能量代谢、突触组分和囊泡运输和再循环。这些发现表明，A β寡聚体改变了海马和大脑皮层中某些蛋白质和磷蛋白的水平，这可能为AD的治疗开辟新的治疗途径。(C)2012爱思唯尔有限公司保留所有权利。
Amyloid beta (A beta) oligomers are presumed to be one of the causes of Alzheimer's disease (AD). Previously, we identified the E693 Delta mutation in amyloid precursor protein (APP) in patients with AD who displayed almost no signals of amyloid plaques in amyloid imaging. We generated APP-transgenic mice expressing the E693 Delta mutation and found that they possessed abundant A beta oligomers from 8 months of age but no amyloid plaques even at 24 months of age, indicating that these mice are a good model to study pathological effects of A beta oligomers. To elucidate whether A beta oligomers affect proteome levels in the brain, we examined the proteins and phosphoproteins for which levels were altered in 12-month-old APP(E693 Delta)-transgenic mice compared with age-matched non-transgenic littermates. By two-dimensional gel electrophoresis (2DE) followed by staining with SYPRO Ruby and Pro-Q Diamond and subsequent mass spectrometry techniques, we identified 17 proteins and 3 phosphoproteins to be significantly changed in the hippocampus and cerebral cortex of APP(E693 Delta)-transgenic mice. Coactosin like-protein, SH3 domain-bind glutamic acid-rich-like protein 3 and astrocytic phosphoprotein PEA-15 isoform 2 were decreased to levels less than 0.6 times those of non-transgenic littermates, whereas dynamin, profilin-2, vacuolar adenosine triphosphatase and creatine kinase B were increased to levels more than 1.5 times those of non-transgenic littermates. Furthermore, 2DE Western Blotting validated the changed levels of dynamin, dihydropyrimidinase-related protein 2 (Dpysl2), and coactosin in APP(E693 Delta)-transgenic mice. Glyoxalase and isocitrate dehydrogenase were increased to levels more than 1.5 times those of non-transgenic littermates. The identified proteins could be classified into several groups that are involved in regulation of different cellular functions, such as cytoskeletal and their interacting proteins, energy metabolism, synaptic component, and vesicle transport and recycling. These findings indicate that A beta oligomers altered the levels of some proteins and phosphoproteins in the hippocampus and cerebral cortex, which could illuminate novel therapeutic avenues for the treatment of AD. (C) 2012 Elsevier Ltd. All rights reserved.