Role of Mitochondrial DNA Mutations in Aging in Neuronal Cells

线粒体 DNA 突变在神经细胞衰老中的作用

• 批准号: 7371075
• 负责人: Yidong Bai
• 金额: $ 28.68万
• 依托单位: UNIVERSITY OF TEXAS HLTH SCIENCE CENTER
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-03-01 至 2011-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7371075
• 关键词: Age; Aging; Aging-Related Process; Apoptosis; Applications Grants; Biochemical; Biogenesis; Brain; Brain region; Cell Culture System; Cell Line; Cell model; Cell physiology; Cells; DNA; Electron Transport; Energy Supply; Evaluation; Exhibits; Functional disorder; Gene Mutation; Generations; Genes; Genetic; Genetic Transcription; Goals; Individual; Lipids; Measures; Methods; Mitochondria; Mitochondrial DNA; Molecular; Molecular Genetics; Monitor; Mus; Mutation; Nerve Endings; Neuroblastoma; Neurons; Nuclear; Oxidative Phosphorylation; Oxidative Stress; Phenotype; Physiological; Proteins; Reactive Oxygen Species; Regulation; Relative (related person); Reporting; Respiration; Role; Sequence Analysis; Signal Transduction; Synaptosomes; System; Techniques; Technology; Testing; Time; Tissues; Translations; age group; age related; established cell line; human tissue; improved; insight; middle age; mitochondrial DNA mutation; mitochondrial dysfunction; mitochondrial genome; mutant; programs; respiratory; theories

Our long-term goal is to study the role of mitochondria in aging. The overall goal of this grant application is to test the mitochondria! theror/ of aging and, in particular, to investigate the role of mitochondrial DNA(mtDNA) mutations in mouse brain during aging. As one of the most favored hypotheses, the mitochondrial theory of aging, predicts that somatic mitochondria DNA (mtDNA) mutations accumulate with time, and the compromised mitochondrial function resulting from these mutations is then responsible for various aging phenotypes. However, there has been no comprehensive study of the overall accumulation of mtDNA mutations during aging, and the physiological consequences of aging-related mtDNA mutations are largely unclear. We recently established a method to transfer mtDNA from mouse nerve endings, i.e., synaptosomes to a cell system, and we have improved methods to isolate and characterize mtDNA mutations. Combined with established mitochondrial molecular genetic and biochemical technologies, we can, for the first time, investigate the accumulation of mtDNA mutations in neuronal cells during aging by evaluating individual mtDNA from the synaptosomes.The particular hypothesis to be tested in this proposal is that mutations in the mitochondrial genome accumulate during aging in mouse neuronal cells; these mtDNA mutations in turn compromise mitochondrial function and may result in oxidative damage to various cellular components including mtDNA in neuronal cells. We will perform genetic and functional analyses of aging-related mtDNA mutations by transferring synaptosomal mtDNA to a cell line system. This will reveal low frequncy mtDNA mutations that could not be identified by other methods. By generating neuronal cell models that carry aging-related mtDNA mutations, we will also charaterize the physiological consequences of the mutations, in particular those related to oxidative damage. The successful completion of this project will not only help to test the mitochondrial theory of aging, but could also provide insights into the underlying mechanisms of the aging process.

我们的长期目标是研究线粒体在衰老中的作用。该赠款申请的总体目标是 测试线粒体！衰老/尤其是研究线粒体DNA（mtDNA）的作用 衰老过程中小鼠大脑中的突变。作为最喜欢的假设之一，线粒体理论 衰老，预测体细分线粒体DNA（mtDNA）突变会随着时间而积聚，而 这些突变导致的线粒体功能受损是各种衰老的原因 表型。但是，尚未对mtDNA的总体积累进行全面研究 衰老过程中的突变以及与衰老相关的mtDNA突变的生理后果很大程度上是 不清楚。我们最近建立了一种从小鼠神经末端转移mtDNA的方法，即突触体 到一个细胞系统，我们有了改进的分离和表征mtDNA突变的方法。与 建立的线粒体分子遗传和生化技术，我们首次可以 通过评估个体，研究神经元细胞中mtDNA突变的积累 来自突触体的mtDNA。在此提案中要检验的特定假设是 线粒体基因组在小鼠神经元细胞衰老过程中积累；这些mtDNA突变反过来 折衷的线粒体功能，可能会对各种细胞成分造成氧化损伤 在神经元细胞中包括mtDNA。我们将对与衰老相关的mtDNA进行遗传和功能分析 通过将突触体mtDNA转移到细胞系系统中来突变。这将揭示低频率mtDNA 无法通过其他方法识别的突变。通过生成携带的神经元细胞模型 与衰老相关的mtDNA突变，我们还将为突变的生理后果而进行展示 特别是与氧化损伤有关的。该项目的成功完成不仅将有助于测试 衰老的线粒体理论，但也可以提供对衰老的潜在机制的见解 过程。