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: 6944710
• 负责人: TERESA G HASTINGS
• 金额: $ 35.09万
• 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 2008-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6944710
• 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（改编自申请人提供的摘要）： 越来越明显的是，线粒体在广泛的 一系列急性和慢性神经退行性疾病。最近的研究表明， 实验室和许多其他人都认为线粒体可能是 杀死神经元的信号也许这个信号的最佳候选人是 线粒体产生活性氧（ROS）。的机制 然而，对脑线粒体的ROS生成知之甚少。此外，委员会认为， 对ROS信号的调节知之甚少。我们建议 研究脑线粒体产生ROS的特点， 培养的神经元在具体目标1中，我们将确定 电子传递链（ETC）是超氧化物的来源，并将 确定ETC抑制离体脑中ROS产生的结果 线粒体因此，我们将能够基于以下模型来模拟神经退行性疾病： 先前关于ETC抑制在帕金森病等疾病中的报道， 老年痴呆症在具体目标2中，我们将研究 调节ROS的产生，并将重点放在改变线粒体的代理人， 膜电位和从ATP解偶联氧消耗的试剂 合成.这些试剂包括钙和解偶联蛋白质的底物。 在具体目标3中，我们将使用一系列神经元制备物来建立 线粒体在其完整细胞中产生ROS的特性 环境我们将使用标准神经元培养物、透化神经元和 急性解离的细胞，沿着ROS的荧光成像，以建立 线粒体ROS产生的神经元特异性机制。在决赛中 具体目的我们将研究是否逆行运输的 线粒体的神经元在体内被改变的氧化损伤。这本小说 这种方法将使我们能够研究细胞线粒体的特性， 稳态，这将提供一个长期的观点，线粒体的作用， 神经变性这些研究将阐明 线粒体和ROS信号在神经元损伤，并将提供新的 在广泛的神经退行性疾病的干预目标。

项目成果

The role of external and matrix pH in mitochondrial reactive oxygen species generation.

• DOI: 10.1074/jbc.m801019200
• 发表时间: 2008-10-24
• 期刊: The Journal of biological chemistry
• 作者: Selivanov VA;Zeak JA;Roca J;Cascante M;Trucco M;Votyakova TV
• 通讯作者: Votyakova TV

Bistability of mitochondrial respiration underlies paradoxical reactive oxygen species generation induced by anoxia.

• DOI: 10.1371/journal.pcbi.1000619
• 发表时间: 2009-12
• 期刊: PLoS computational biology
• 影响因子: 4.3
• 作者: Selivanov VA;Votyakova TV;Zeak JA;Trucco M;Roca J;Cascante M
• 通讯作者: Cascante M

Reactive oxygen species production by forward and reverse electron fluxes in the mitochondrial respiratory chain.

• DOI: 10.1371/journal.pcbi.1001115
• 发表时间: 2011-03
• 期刊: PLoS computational biology
• 影响因子: 4.3
• 作者: Selivanov VA;Votyakova TV;Pivtoraiko VN;Zeak J;Sukhomlin T;Trucco M;Roca J;Cascante M
• 通讯作者: Cascante M

Spontaneous changes in mitochondrial membrane potential in single isolated brain mitochondria.

• DOI: 10.1016/s0006-3495(03)74755-9
• 发表时间: 2003-11
• 期刊: Biophysical journal
• 影响因子: 3.4
• 作者: O. Vergun;T. Votyakova;I. Reynolds