Alcohol And Mitochondrial Redistribution in Neurons

酒精和神经元中的线粒体重新分布

• 批准号: 7065205
• 负责人: SHIVANI KAUSHAL MAFFI
• 金额: $ 16.93万
• 依托单位: UNIVERSITY OF TEXAS HLTH SCIENCE CENTER
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-05-12 至 2008-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7065205
• 关键词: aldehydes; axon; cerebral cortex; confocal scanning microscopy; dendrites; disease /disorder model; ethanol; fetal alcohol syndrome; glutathione; immunofluorescence technique; intracellular transport; laboratory rat; microtubules; mitochondria; mitochondrial DNA; mitochondrial disease /disorder; molecular /cellular imaging; neurogenesis; neurons; neurotoxicology; oxidative stress; tissue /cell culture

DESCRIPTION (provided by applicant): Alcohol use/abuse is a leading cause of neurological damage in humans, especially in the context of alcohol damage to the developing nervous system in fetuses. However, the underlying mechanisms of this neurotoxicity are poorly understood. With this in mind, my goal is to understand alcohol-induced alterations in mitochondrial movements in neurons. The strategic positioning of mitochondria along neuronal processes plays a critical/central role in energy production and Ca2+ homeostasis. Moreover, the maintenance of structurally and functionally distinct axonal and dendritic processes is important for normal neuronal function. This proposal will address a new mechanism by which ethanol may compromise neuron function at the mitochondrial level. Alcohol may alter mitochondrial redistribution in neuronal processes via a rapid onset of oxidative stress (OS), and disrupting the cytoskeletal proteins. One or both of these factors may affect mitochondrial motility. Hence, I hypothesize that alcohol-induced oxidative stress results in the disruption of the temporal movement and spatial distribution of mitochondria in the neuronal processes, and this may contribute to neuronal dysfunction or death. To test this, we will capitalize on a novel live cell imaging approach in conjunction with low density seeded of primary cortical neuron cultures. Our preliminary results provide a strong rationale for our hypothesis and we propose two specific aims. First, determine the effects of alcohol on mitochondrial motility (dose and time-dependent) i.e. changes in direction and its velocity in the axons and dendrites, This approach will utilize live cell imaging confocal, multiphoton and fluorescence microscopic techniques to visually and quantitatively assess mitochondria undergoing anterograde, retrograde and saltatory movements, and, net and instantaneous velocity; Second, determine using similar approaches, whether potential mediators of alcohol-induced oxidative stress (HNE) influences mitochondrial redistribution. The work on 4-hydroxynonenal (HNE) and its relationship to mitochondrial movements, cytoskeletal disruption, its localization in neurons and glutathione status will provide novel insights into the mechanism of action of ethanol. These studies are significant as mitochondrial diseases are mainly reported in organs that have high energy consumption, such as liver, skeletal tissue and brain. Hence, the information gathered will have implications to our future understanding of the role of mitochondrial motility in homeostasis and disease states.

描述（由申请人提供）：饮酒/滥用是人类神经系统损害的主要原因，尤其是在酒精对胎儿发展中神经系统损害的情况下。但是，这种神经毒性的基本机制知之甚少。考虑到这一点，我的目标是了解酒精引起的神经元线粒体运动的改变。线粒体沿神经元过程的战略定位在能源生产和CA2+稳态中起关键/中心作用。此外，在结构和功能上不同的轴突和树突过程中维持对正常神经元功能很重要。该提案将解决一种新的机制，乙醇可能会在线粒体级别损害神经元功能。酒精可能会通过氧化应激（OS）快速发作并破坏细胞骨架蛋白的神经元过程中的线粒体重新分布。这些因素中的一个或两个可能影响线粒体运动。因此，我假设酒精诱导的氧化应激会导致神经元过程中线粒体的时间运动和空间分布的破坏，这可能导致神经元功能障碍或死亡。为了测试这一点，我们将利用一种新型的活细胞成像方法，并结合原发性皮质神经元培养物的低密度种子。我们的初步结果为我们的假设提供了有力的理由，我们提出了两个具体的目标。 First, determine the effects of alcohol on mitochondrial motility (dose and time-dependent) i.e. changes in direction and its velocity in the axons and dendrites, This approach will utilize live cell imaging confocal, multiphoton and fluorescence microscopic techniques to visually and quantitatively assess mitochondria undergoing anterograde, retrograde and saltatory movements, and, net and instantaneous velocity;其次，使用类似方法确定酒精诱导的氧化应激（HNE）的潜在介体是否影响线粒体重新分布。关于4-羟基诺纳尔（HNE）的工作及其与线粒体运动，细胞骨架破坏，其在神经元中的定位和谷胱甘肽地位的关系，将为乙醇作用机理提供新的见解。这些研究很重要，因为线粒体疾病主要报道在肝脏，骨骼组织和大脑等能量较高的器官中。因此，收集的信息将对我们对线粒体运动在体内平衡和 疾病状态。