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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：
7475679
负责人：
PHILIP W. LANDFIELD
金额：
$ 30.98万
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/7475679
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.

β-淀粉样肽 (Abeta) 是阿尔茨海默病 (AD) 发病机制的核心，而 AD 大脑处于广泛的氧化应激之下。我们实验室之前的研究将这两个观察结果结合成 AD 神经变性模型，该模型基于 Abeta 相关的自由基氧化应激。在神经元中，Abeta 相关的自由基氧化应激和毒性主要取决于 Abeta(1-42) 的蛋氨酸残基 35，并且两者都可以通过外源抗氧化剂预防或调节。我们使用新兴的蛋白质组学技术，结合免疫印迹方法，来识别 AD 大脑中特异性氧化修饰的蛋白质。我们现在建议将这些观察扩展到散发性和遗传性 AD 及其啮齿动物模型，以深入了解 Abeta (1-42) 在该肽的氧化应激和神经毒性特性中的机制。在具体目标#1中，我们将使用蛋白质组学来检验以下假设：遗传性和散发性 AD 中存在一组与正常衰老不同的常见氧化蛋白，并且这些共同的氧化蛋白在 AD 的发病机制中很重要。假设在遗传性 AD 小鼠模型中进行的类似研究能够确定相同的常见蛋白质。在特定目标 #2 中，我们将测试以下假设：与野生型小鼠的成年大脑相比，在特定目标 #1 中选择的模型中的成年大脑线粒体中会发现与 AD 相关的基因突变导致的线粒体功能障碍和大脑中 Ca 2+ 积累。在研究系统中通过蛋白质组学鉴定的那些蛋白质将在 SY5Y 细胞中表达，并用 Abeta(1-42) 进行攻击，以确定是否对氧化应激、线粒体功能障碍、Ca 2+ 积累和细胞毒性的脆弱性增加。在特定目标 #3 中，我们将使用蛋白质组学来检验以下假设：在从特定目标 #1 中选择的家族性 AD 啮齿动物模型获得的胚胎神经元培养物中，Abeta(1-42) 诱导的氧化应激会导致与该动物的成年大脑和人类家族性 AD 大脑中存在的相同氧化修饰蛋白质。在具体目标#4中，我们将研究氧化应激是否会诱导线粒体从特定目标 #1 中选择的遗传性 AD 啮齿动物模型获得的神经元培养物的改变，或者这些培养物中诱导的线粒体改变是否会导致氧化应激。在具体目标 # 5 中，我们将测试以下假设：在具体目标 # 1 中选择的啮齿动物模型中，内源性或外源性抗氧化剂可保护大脑体内免受氧化应激。这是一套全面的拟议研究，在系统中采用新颖的方法，从散发性和遗传性 AD 大脑到 AD 遗传动物模型，再到神经元培养，旨在深入了解 Abeta 相关的自由基氧化应激和神经毒性及其通过内源性和外源性抗氧化剂的调节。这些研究将加深对这一影响数百万美国人的重要痴呆症的潜在治疗干预的了解。