Proteomics of age-related neurodegeneration

年龄相关神经变性的蛋白质组学

• 批准号: 6950253
• 负责人: AUSTIN YANG
• 金额: $ 32.52万
• 依托单位: UNIVERSITY OF SOUTHERN CALIFORNIA
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-09-30 至 2006-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6950253
• 关键词: Alzheimer' s disease; affinity chromatography; aging; amyloid proteins; bioinformatics; biological signal transduction; biotechnology; clinical research; dietary restriction; free radical oxygen; genetically modified animals; high throughput technology; laboratory mouse; liquid chromatography mass spectrometry; neural degeneration; neuropathology; neurophysiology; nitration; oxidative stress; phosphorylation; posttranslational modifications; presenilin; proteomics; statistics /biometry; two dimensional gel electrophoresis

DESCRIPTION (provided by applicant): We will use a newly developed high throughput sensitive, quantitative proteomic approach to detect key proteins and signaling pathways that are oxidatively damaged during aging and Alzheimer's disease (AD). We will accurately identify the initial targets of age- and ABeta-mediated oxidative stress and their association with neuropathology. This addresses several key elements of this RFA as both normal brain aging and AD are associated with free radical-mediated damage and a loss of synaptic function. which may be a primary cause of cognitive impairments. Many age-related oxidative stress-mediated processes are attenuated by Caloric Restriction (CR). We have recently demonstarted that CR also reduces ABeta deposition and glial activation in the bigenic mutated presenilin & amyloid precursor protein (PS1+APP) mouse models of AD. In addition, using liquid chromatography-mass spectrmetry proteomic methodology, we discovered that certain proteins associated with learning and memory (e.g., NMDA/AMPA receptor, fyn, CaMKII) are oxidatively damaged in the PS1+APP mouse. Therefore, we propose to identify key proteins and signaling pathways involved in free radical mediated syanptic dysfuntion during normal aging and AD using our sensitive and accurate proteomic approach in the PS1+APP mouse. Since CR attenuates many age-related oxidative stress mediated processes, we will determine which proteins and pathways are protected by this dietary intervention. 1: We will use quantitative proteomics to determine the initial targets of free radicals in PSI+APP transgenic and nontransgenic mice to establish the functional links between neuropathological changes and oxidative modification of key synaptic signaling proteins. 2: We will identify the phosphorylation alterations in cell signaling pathways that mediate synaptic dysfunction during aging and Abeta-mediated pathology. 3: We will perform statistical validation of protein identifications from database searches and we will use bioinformatcis to integrate our proteomic data into signaling pathways to identify the underlying mechanisms involved in synaptic dysfunction in the PS1+APP mouse model.

描述（由申请人提供）：我们将使用新开发的高吞吐量敏感的定量蛋白质组学方法来检测在衰老和阿尔茨海默氏病（AD）期间氧化受损的关键蛋白质和信号通路。我们将准确地确定年龄和ABETA介导的氧化应激的初始靶标及其与神经病理学的关联。这解决了该RFA的几个关键要素，因为正常的脑老化和AD都与自由基介导的损伤和突触功能丧失有关。这可能是认知障碍的主要原因。许多与年龄相关的氧化应激介导的过程通过热量限制（CR）减弱。最近，我们推出了CR还减少了AD的脂质突变的Presenilin和淀粉样蛋白前体蛋白（PS1+APP）AD的AD中的ABETA沉积和神经胶质激活。此外，使用液相色谱质谱蛋白质组学方法，我们发现与学习和记忆相关的某些蛋白质（例如NMDA/AMPA受体，FYN，CAMKII）在PS1+App小鼠中受到氧化损害。因此，我们建议在正常衰老期间鉴定参与自由基介导的syantagic功能障碍的关键蛋白质和信号通路，并使用我们在PS1+App小鼠中使用敏感和准确的蛋白质组学方法进行AD。由于CR会减弱许多与年龄相关的氧化应激介导的过程，因此我们将确定哪些蛋白质和途径受到这种饮食干预的保护。 1：我们将使用定量蛋白质组学来确定PSI+APP转基因和非转基因小鼠中自由基的初始靶标，以建立神经病理学变化与关键突触信号蛋白的氧化修饰之间的功能联系。 2：我们将确定在衰老和ABETA介导的病理过程中介导突触功能障碍的细胞信号通路的磷酸化改变。 3：我们将对来自数据库搜索的蛋白质识别进行统计验证，并将使用Bioinformatcis将蛋白质组学数据整合到信号通路中，以识别PS1+App鼠标模型中突触功能障碍所涉及的潜在机制。