Control of neuronal ROS generation by membrane potential

通过膜电位控制神经元ROS的产生

• 批准号: 6796688
• 负责人: TERESA G HASTINGS
• 金额: $ 41.84万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-09-30 至 2006-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6796688
• 关键词: brain metabolism; calcium metabolism; cell component structure /function; chemical structure function; cytotoxicity; electron transport; enzyme inhibitors; fluorescent dye /probe; free radical oxygen; glutamates; laboratory rat; membrane potentials; mitochondria; neural degeneration; neuronal transport; neurons; neuropathology; neurophysiology; neuroregulation; oxidative stress; tissue /cell culture

DESCRIPTION (Adapted from the abstract provided by the applicant): It is becoming increasingly apparent that mitochondria play a critical role in a wide range of acute and chronic neurodegenerative diseases. Recent studies from this laboratory and many others have suggested that mitochondria may be the source of a signal that kills neurons. Perhaps the best candidate for this signal is mitochondrial generation of reactive oxygen species (ROS). The mechanisms of ROS generation by brain mitochondria, however, are poorly understood. Moreover, little is known about the regulation of the ROS signal. We propose to investigate the characteristics of ROS generation by brain mitochondria and cultured neurons. In Specific Aim 1 we will determine the site(s) in the electron transport chain (ETC) that are the source of superoxide, and will determine the consequence of ETC inhibition on ROS generation in isolated brain mitochondria. Thus we will be able to model neurodegenerative disease based on prior reports of ETC inhibition in disorders such as Parkinson's disease and Alzheimer's disease. In Specific Aim 2 we will investigate the mechanisms that regulate ROS generation and will focus on agents that alter the mitochondrial membrane potential and agents that uncouple oxygen consumption from ATP synthesis. These agents include calcium and substrates for uncoupling proteins. In Specific Aim 3 we will use a series of neuron preparations to establish the properties of ROS generation by mitochondria in their intact cellular environment. We will use standard neuronal cultures, permeabilized neurons and acutely dissociated cells, along with fluorescence imaging of ROS, to establish neuron-specific mechanisms of mitochondrial ROS generation. In the final Specific Aim we will investigate whether the retrograde transport of mitochondria by neurons in vivo is altered by oxidative injury. This novel approach will allow us to investigate the properties of cellular mitochondrial homeostasis which will provide a long-term view of the role of mitochondria in neurodegeneration. These studies will illuminate a critical role of mitochondria and ROS signaling in neuronal injury, and will provide novel targets for intervention in a wide range of neurodegenerative disease.

描述（根据申请人提供的摘要改编）： 越来越明显的是，线粒体在广泛的 急性和慢性神经退行性疾病的范围。最近的研究 实验室和其他许多人建议线粒体可能是来源 杀死神经元的信号。也许这个信号的最佳候选者是 线粒体生成活性氧（ROS）。机制 然而，脑线粒体产生ROS的人数很少。而且， 关于ROS信号的调节知之甚少。我们建议 研究脑线粒体和ROS产生的特征 培养的神经元。在特定目标1中，我们将确定位置 电子传输链（ETC）是超氧化物的来源，将 确定ETC抑制对孤立大脑中ROS产生的结果 线粒体。因此，我们将能够基于 事先报道了帕金森氏病和 阿尔茨海默氏病。在特定目的2中，我们将研究的机制 调节ROS的产生，并将专注于改变线粒体的代理 膜电位和脱离ATP消耗氧气的试剂 合成。这些药物包括钙和底物的解偶联蛋白。 在特定目标3中，我们将使用一系列神经元制剂来确定 线粒体在完整细胞中产生ROS的特性 环境。我们将使用标准的神经元培养物，透化神经元和 急性解离的细胞以及ROS的荧光成像，建立 线粒体ROS产生的神经元特异性机制。在决赛中 具体目的我们将调查是否逆行运输 神经元在体内的线粒体会因氧化损伤而改变。这本小说 方法将使我们能够研究细胞线粒体的特性 体内平衡将长期观察线粒体在 神经变性。这些研究将阐明 神经元损伤中的线粒体和ROS信号传导，并将提供新颖 干预多种神经退行性疾病的目标。