Aging-Related Models and Mitochondrial Bioenergetics

衰老相关模型和线粒体生物能学

• 批准号: 6866416
• 负责人: David Nicholls
• 金额: $ 36.86万
• 依托单位: BUCK INSTITUTE FOR RESEARCH ON AGING
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-03-01 至 2007-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6866416
• 关键词: action potentials; aging; bioenergetics; biological models; calcium ion; cellular respiration; cerebellum; free radical oxygen; functional /structural genomics; gene targeting; genetically modified animals; glutamates; hippocampus; laboratory mouse; membrane potentials; mitochondria; neurons; neurotoxins; oxidative stress; plant insecticide; proteomics; superoxide dismutase; tetany; tissue /cell culture

DESCRIPTION (provided by applicant): Oxidative stress and decreased mitochondrial respiratory chain capacity are both implicated in normal aging and may contribute to the increased incidence of neurodegenerative disease. It is possible that the slow insidious onset of neurodegenerative disease is because neuronal damage may advance in a quantal manner, each downward step being initiated by an acute energy deficit. The aim of this research proposal is to investigate the consequences of such bioenergetic compromise upon the ability of primary neuronal cultures from mouse hippocampus and cerebellum to withstand the patho-physiological stresses of acute glutamate excitotoxicity and tetanic action potential firing. In order to model enhanced oxidative stress, cells will be cultured from mice heterozygous for the Sod2 gene encoding mitochondrial Mn-superoxide dismutase. Chronic complex I restriction will be modelled by culturing cells from the wild-type mice in the presence of low concentrations of the respiratory chain inhibitor rotenone. The central bioenergetic parameter to be monitored will be the mitochondrial membrane potential, membrane potential while the key cellular functions controlled by membrane potential namely ATP synthesis, Ca2+ sequestration and the generation and detoxification of reactive oxygen species (ROS), will be monitored in parallel. Oxidative stress derived from Ca2+loaded mitochondria is implicated in the excitotoxic cell death induced by chronic NMDA receptor activation. Since ROS generation by isolated mitochondria is highly dependent upon membrane potential the hypothesis will be investigated that endogenous or synthetic means of controlling membrane potential can alleviate the deleterious consequences to the neurons of these bioenergetic stresses. The role of the novel candidate uncoupling protein UCP5 in controlling membrane potential will be assessed by in vitro over-expression, while the efficacy of synthetic protophores and lipophilic cations will be investigated. The hypothesis that this project will test that fine control of membrane potential by endogenous or artificial means may be a mechanism to counteract any increase, resulting from age related mitochondrial function, in the susceptabilty of neurons to excititoxic or epileptiform stress. Bioenergetic, proteomic and genomic information will be integrated in this study. The long-term goals of this project are 1. To facilitate the design of more precise mouse models of aging. 2. To establish the role of mitochondria in normal aging and aging-related neurodegenrative disorder. 3. To indicate therapeutic strategies whereby the deleterious consequences of aging-related changes in mitochondrial function can be minimized.

描述（由申请人提供）：氧化应激和减少 线粒体呼吸链能力都与正常衰老有关 并可能导致神经退行性疾病的发病率增加。它 神经退行性疾病的慢阴险发作可能是 由于神经元损害可能以数量的方式前进，所以每个下降步骤 由急性能量不足引发。该研究建议的目的 是为了研究这种生物能妥协对 小鼠海马和小脑的原发性神经元培养的能力 承受急性谷氨酸兴奋性毒性的病理生理应力 和跨动作潜在的射击。为了建模增强的氧化剂 压力，细胞将从小鼠的杂合子中培养为SOD2基因 编码线粒体Mn-塞氧化物歧化酶。慢性综合体I限制 将通过在存在下从野生型小鼠中培养细胞来建模 低浓度的呼吸链抑制剂烤面包酮。中央 要监测的生物能参数将是线粒体膜 电势，膜电位，而关键细胞功能由 膜电势，即ATP合成，Ca2+隔离和产生 将监测活性氧（ROS）的排毒 平行线。与Ca2+负载线粒体衍生的氧化应激有关 在慢性NMDA受体激活引起的兴奋性细胞死亡中。 由于通过孤立线粒体产生ROS，高度依赖 膜电位该假设将研究内源性或 控制膜电位的合成手段可以减轻 这些生物能力的神经元的有害后果。这 新型候选蛋白UCP5在控制膜中的作用 潜力将通过体外过度表达来评估，而的功效 将研究合成的原始阳离子和亲脂阳离子。 该项目将测试膜的精细控制的假设 通过内源或人工手段的潜力可能是抵消的机制 在与年龄相关的线粒体功能中导致的任何增加 神经元的敏感性或癫痫样应激的敏感性。生物元素， 蛋白质组学和基因组信息将集成到本研究中。长期 该项目的目标是1。促进更精确的设计 衰老的小鼠模型。 2。确定线粒体在正常中的作用 衰老和与衰老有关的神经退行性疾病。 3。表示治疗 策略，与衰老相关的变化的有害后果 线粒体功能可以最小化。