Seahorse XF-96 Analyzer for Non-Invasive Monitoring of Mitochondrial Function

描述（由申请人提供）：线粒体在许多神经系统疾病的发病机制中起关键作用，部分原因是线粒体在突触处和沿着轴突的神经元活动中的独特功能。已经开发了利用原代神经元培养物、外植体/切片培养物以及神经元和神经胶质细胞系的神经退行性疾病的体外模型，以研究疾病机制并建立用于药物筛选的测定。线粒体在氧化应激、轴突变性、细胞死亡和神经元能量代谢中的作用正在这些体外模型中进行研究，但目前大多数可用的测定方法都有局限性。许多关于线粒体功能的测定需要从细胞中分离线粒体，从而消除细胞内信号传导对线粒体功能的贡献。其他测定是间接的，并且观察线粒体功能障碍的后果，并且通常是静态测定。一种在神经元和神经胶质细胞内实时评估线粒体功能的仪器对于检验线粒体在神经退行性疾病发病机制中的相关性的假设具有重要价值。Seahorse XF-96分析仪就是这样一种仪器，它允许人们实时评估活细胞中的耗氧率（OCR）和细胞外酸化率（ECAR，即质子产生）。XF-96的主要研究者和主要使用者都是研究神经退行性疾病机制的神经学家和神经科学家，包括运动神经元疾病、多发性硬化症、人类免疫缺陷病毒（HIV）痴呆和周围神经病。正如提案中所概述的那样，XF-96分析仪将允许研究人员随着时间的推移测量整个神经元和神经胶质细胞群体中的线粒体功能，从而消除了在不同时间点使用多个样本进行定时实验的需要;从而加快了研究并促进了新科学方向的探索。 公共卫生相关性：拟议购买Seahorse XF 96分析仪将有助于加快正在进行的旨在了解各种神经退行性疾病机制的研究，包括痴呆症，运动神经元疾病，多发性硬化症和周围神经病变。这些神经退行性疾病是影响大部分人口的最常见和最具破坏性的神经系统疾病。

DESCRIPTION (provided by applicant): Mitochondria play a critical role in pathogenesis of many neurological disorders, partly because of the unique function of mitochondria in neuronal activity at the synapses and along the axons. In vitro models of neurodegenerative disorders utilizing primary neuronal cultures, explants/slice cultures and neuronal and glial cell lines have been developed to study disease mechanisms and establish assays for drug screening. Role of mitochondria in oxidative stress, axonal degeneration, cell death and neuronal energy metabolism are being studied in these in vitro models, but most of the currently available assays have limitations. Many of the assays on mitochondrial function require isolation of mitochondria from the cells eliminating contribution of intracellular signaling on mitochondrial function. Other assays are indirect and look at consequences of mitochondrial dysfunction and are often static assays. An instrument that evaluates mitochondrial function in real-time within neuronal and glial cells would be of great value to test hypotheses on the relevance of mitochondria in neurodegenerative disease pathogenesis. The Seahorse XF-96 analyzer is such an instrument allowing one to evaluate oxygen consumption rate (OCR) and extracellular acidification rate (ECAR, i.e. proton production) in living cells in real-time. The principal investigator and major users of the proposed XF-96 are all neurologists and neuroscientists studying neurodegenerative disease mechanisms, including motor neuron disease, multiple sclerosis, human immunodeficiency virus (HIV) dementia, and peripheral neuropathies. As outlined in the proposal, the XF-96 analyzer will allow the investigators to measure mitochondrial function in whole neuronal and glial cell populations over time eliminating the need for timed experiments with multiple samples at different time points; thus expediting the research and facilitating explorations of new scientific directions. PUBLIC HEALTH RELEVANCE: The proposed purchase of Seahorse XF96 analyzer will help expedite ongoing studies aimed at understanding the mechanisms of a variety of neurodegenerative diseases, including dementias, motor neuron disease, multiple sclerosis and peripheral neuropathies. These neurodegenerative diseases are among the most common and devastating types of neurological disorders affecting a large segment of the population.