Ferreira I Luísa, Carmo Catarina, Naia Luana, I Mota Sandra, Cristina Rego A
CNC-Center for Neuroscience and Cell Biology, University of Coimbra-polo I, Coimbra, Portugal.
IIIUC-Institute for Interdisciplinary Research, University of Coimbra-polo II, Coimbra, Portugal.
Methods Mol Biol. 2018;1780:415-442. doi: 10.1007/978-1-4939-7825-0_19.
Mitochondrial dysfunction has gained a preponderant role in the pathogenesis of Huntington's disease (HD). Mutant huntingtin (mHTT) directly interacts with mitochondria in a deleterious manner. As the central hub of the cell, not only mitochondrial bioenergetics is affected but there is also diminished mitochondrial membrane potential (Δψ ) and altered production of reactive oxygen species (ROS). Restoration of mitochondrial function has proven to be a major player in the search and establishment of therapeutics for HD patients. As such, performing an overall study of mitochondrial function is crucial. In this chapter, we describe some methodologies used to study mitochondrial function by determining the oxygen consumption, changes in Δψ , mitochondrial calcium handling, and levels of mitochondrial ROS. Here we focus on biological samples derived from HD versus control cells and/or animal models, namely functional isolated brain mitochondria, an ex vivo animal model, and cultured cells, including cell lines and primary neural cultures, as in vitro models.
线粒体功能障碍在亨廷顿舞蹈病（HD）的发病机制中发挥着重要作用。突变型亨廷顿蛋白（mHTT）以有害方式直接与线粒体相互作用。作为细胞的核心枢纽，不仅线粒体生物能量学受到影响，线粒体膜电位（Δψ）也会降低，活性氧（ROS）的产生也会改变。事实证明，恢复线粒体功能是寻找和确立HD患者治疗方法的关键因素。因此，对线粒体功能进行全面研究至关重要。在本章中，我们描述了一些用于研究线粒体功能的方法，这些方法通过测定氧气消耗、Δψ的变化、线粒体钙处理以及线粒体ROS水平来实现。在这里，我们重点关注来自HD与对照细胞和/或动物模型的生物样本，即功能分离的脑线粒体、离体动物模型和培养细胞，包括细胞系和原代神经培养物，作为体外模型。
