CA2+ REGULATION AND MITOCHONDRIA IN BRAIN AGING AND ALZHEIMER'S DISEASE

脑老化和阿尔茨海默病中的 CA2 调节和线粒体

基本信息

批准号：
7674568
负责人：
PHILIP W. LANDFIELD
金额：
$ 36.66万
依托单位：
UNIVERSITY OF KENTUCKY
依托单位国家：
美国
项目类别：
财政年份：
资助国家：
美国
起止时间：
至
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/7674568
关键词：
Adult Affect Age Age-Months Aging Agonist Algorithms Alzheimer&apos s Disease Alzheimer&apos s disease model American Amyloid Amyloid beta-Protein Animal Model Animals Antioxidants Astrocytes Behavioral Binding Binding Proteins Biochemical Bioinformatics Brain Caffeine Calcineurin Calcium Cell Aging Cells Collaborations Complement Complex Coupled Cultured Cells Data Deposition Disease Embryo Endoplasmic Reticulum Estrogen Receptors Event Exhibits Family Free Radicals Frequencies Gene Expression Gene Expression Process Gene Expression Profiling Gene Mutation Gene Transfer Generations Genes Genetic Genomics Glial Fibrillary Acidic Protein Glutamates Goals Green Fluorescent Proteins Growth Factor Hippocampus (Brain)Human Image Impaired cognition Inflammatory Inherited Intervention Laboratories Lead Learning Mediating Membrane Methionine Methods Microarray Analysis Mitochondria Modeling Molecular Mus Myelin Proteins Nerve Degeneration Neuroglia Neurons Nimodipine Outcome Study Oxidative Stress Pathogenesis Pathology Pathway interactions Pattern Peptides Phase Physiological Polymerase Chain Reaction Positioning Attribute Preparation Procedures Process Progress Reports Property Protein Overexpression Proteins Proteomics Range Rattus Regulation Regulatory Pathway Research Resources Rodent Model Running Signal Transduction Slice Small Interfering RNA Source Specificity Steroids Synapses System Techniques Testing Therapeutic Intervention Time Toxic effect Transgenic Mice Transgenic Organisms Up-Regulation Ursidae Family Validation Viral Wild Type Mouse Work age related aged aging brain base brain cell cell age cell type cytotoxicity density design familial Alzheimer disease in vivo inhibitor/antagonist insight interdisciplinary approach knock-down mature animal mitochondrial dysfunction mouse model multidisciplinary mutant neurotoxic neurotoxicity normal aging novel strategies prevent receptor receptor expression research study response steroid hormone steroid hormone receptor tool transgenic model of alzheimer disease voltage

项目摘要

Amyloid beta-peptide (Abeta) is central to the pathogenesis of Alzheimer's disease (AD), and the AD brain is under extensive oxidative stress. Previous research from our laboratory combined these two observations into a model for neurodegeneration in AD, a model based on Abeta-associated free radical oxidative stress. In neurons, Abeta-associated free radical oxidative stress and toxicity depend critically on methionine residue 35 of Abeta(1-42), and both are prevented or modulated by exogenous antioxidants. We have used the emerging techniques of proteomics, coupled with immunoblottmg methods, to identify specifically oxidatively modified proteins in AD brain. We now propose to extend these observations to sporadic and inherited AD and rodent models thereof to gain insight into the mechanisms of Abeta (1-42) in the oxidative stress and neurotoxic properties of this peptide. In Specific Aim # 1, we will use proteomics to test the hypothesis that there will be a common set of oxidized proteins in inherited and sporadic AD that differ from those of normal aging, and that these oxidized proteins in common are important in the pathogenesis of AD. Similar studies in mice models of inherited AD are hypothesized to lead to the identity of the same common proteins. In Specific Aim #2, we will test the hypothesis that mitochondrial dysfunction and Ca 2+ accumulation in brain from genetic mutations relevant to AD will be found in adult brain mitochondria from the model chosen in Specific Aim #1 compared to adult brain from wild-type mice. Those proteins identified by proteomics in the systems studied will be expressed in SY5Y cells and challenged with Abeta(1-42) to determine if there is increased vulnerability to oxidative stress, mitochondrial dysfunction, Ca 2+ accumulation, and cytotoxicity. In Specific Aim #3, we will use proteomics to test the hypothesis that Abeta(1-42)-induced oxidative stress in embryonic neuronal cultures obtained from the rodent model of familial AD chosen in Specific Aim # 1 leads to the same oxidatively modified proteins as exists in adult brain from this animal and in human familial AD brain. In Specific Aim #4, we will investigate whether oxidative stress induces mitochondrial alterations in neuronal cultures obtained from the rodent model of inherited AD chosen in Specific Aim #1, or whether mitochondrial alterations induced in these cultures leads to oxidative stress. In Specific Aim # 5, we will test the hypothesis that endogenous or exogenous antioxidants protect brain in-vivo against oxidative stress in the rodent model chosen in Specific Aim # 1. This is a comprehensive set of proposed studies, employing novel approaches in systems ranging from sporadic and inherited AD brain, to genetic animal models of AD, to neuronal cultures, designed to gain insight into Abeta-associated free radical oxidative stress and neurotoxicity and their modulation by endogenous and exogenous antioxidants. The increased understanding that will result from these studies will provide insight into potential therapeutic interventions in this important dementing disorder that affects millions of Americans.

淀粉样β-肽（ABETA）是阿尔茨海默氏病（AD）发病机理的核心，并且AD脑处于广泛的氧化应激下。我们实验室的先前研究将这两个观察结果结合在一起，成为AD中神经退行性的模型，该模型基于与Abeta相关的自由基氧化应激。在神经元中，与ABETA相关的自由基氧化应激和毒性严重取决于Abeta的蛋氨酸残基35（1-42），并且两者都由外源性抗氧化剂预防或调节。我们已经使用了蛋白质组学的新兴技术，再加上免疫模胞方法，以识别AD脑中特定氧化氧化蛋白的特定氧化蛋白。现在，我们建议将这些观察结果扩展到零星和遗传的AD和啮齿动物模型，以深入了解该肽的氧化应激和神经毒性特性中Abeta（1-42）的机制。在特定的目标＃1中，我们将使用蛋白质组学来检验以下假设：遗传和零星AD中会有一组氧化蛋白与正常衰老不同的蛋白质，并且这些共同的氧化蛋白在AD的发病机构中很重要。假设在遗传AD的小鼠模型中类似的研究导致相同常见蛋白的身份。在特定的目标＃2中，我们将检验以下假设：与野生型小鼠相比，在成人脑线粒体中，在成人脑线粒体中，与AD相关的遗传突变中的线粒体功能障碍和Ca 2+积累。研究系统中蛋白质组学鉴定的蛋白质将在SY5Y细胞中表达，并在ABETA（1-42）中挑战，以确定是否存在增加氧化应激，线粒体功能障碍，Ca 2+积累和细胞毒性的脆弱性。在特定的目标＃3中，我们将使用蛋白质组学来检验以下假设：Abeta（1-42）诱导的氧化应激在胚胎神经元培养物中从特定目标1中选择的啮齿动物模型获得的胚胎神经元培养物中获得的氧化应激会导致相同的氧化性修饰蛋白，因为这种动物和人类家族脑中的成人大脑中存在于成人大脑中，并且是人类家族脑中存在的。在特定的目标＃4中，我们将研究氧化应激是否诱导线粒体从特定目标1中选择的遗传AD的啮齿动物模型获得的神经元培养物的改变，或者在这些培养物中诱导的线粒体改变是否导致氧化应激。 In Specific Aim # 5, we will test the hypothesis that endogenous or exogenous antioxidants protect brain in-vivo against oxidative stress in the rodent model chosen in Specific Aim # 1. This is a comprehensive set of proposed studies, employing novel approaches in systems ranging from sporadic and inherited AD brain, to genetic animal models of AD, to neuronal cultures, designed to gain insight into Abeta-associated free radical oxidative压力和神经毒性及其通过内源性和外源性抗氧化剂的调节。这些研究将产生的越来越多的理解将为影响数百万美国人的这种重要痴呆障碍的潜在治疗干预措施提供见解。