Grad Leslie I, Sayles Leanne C, Lemire Bernard D
Membrane Protein Research Group, Department of Biochemistry, University of Alberta, Edmonton, Canada.
Methods Mol Biol. 2007;372:51-66. doi: 10.1007/978-1-59745-365-3_4.
Mitochondria are essential organelles with central roles in diverse cellular processes such as apoptosis, energy production via oxidative phosphorylation, ion homeostasis, and the synthesis of heme, lipid, amino acids, and iron-sulfur clusters. Defects in the mitochondrial respiratory chain lead to or are associated with a wide variety of diseases in humans. The nematode Caenorhabditis elegans provides a powerful genetic and developmental model system for reproducing deleterious mutations causing mitochondrial dysfunction and investigating their metabolic consequences and their mechanisms of pathology. In this chapter, we describe the growth of C. elegans in liquid culture, the isolation of crude and purified mitochondria, and polarographic and histochemical approaches for measuring mitochondrial respiratory chain function.
线粒体是重要的细胞器,在多种细胞过程中发挥核心作用,如细胞凋亡、通过氧化磷酸化产生能量、离子稳态以及血红素、脂质、氨基酸和铁硫簇的合成。线粒体呼吸链缺陷会导致人类多种疾病或与之相关。线虫秀丽隐杆线虫为再现导致线粒体功能障碍的有害突变、研究其代谢后果及其病理机制提供了一个强大的遗传和发育模型系统。在本章中,我们描述了秀丽隐杆线虫在液体培养中的生长、粗制和纯化线粒体的分离,以及用于测量线粒体呼吸链功能的极谱法和组织化学方法。