Clem Jones Centre for Ageing Dementia Research, Queensland Brain Institute, The University of Queensland, Brisbane, Australia.
Nat Cell Biol. 2018 Mar;20(3):352-360. doi: 10.1038/s41556-017-0023-x. Epub 2018 Jan 22.
Although mitochondria are ubiquitous organelles, they exhibit tissue-specific morphology, dynamics and function. Here, we describe a robust approach to isolate mitochondria from specific cells of diverse tissue systems in Caenorhabditis elegans. Cell-specific mitochondrial affinity purification (CS-MAP) yields intact and functional mitochondria with exceptional purity and sensitivity (>96% enrichment, >96% purity, and single-cell and single-animal resolution), enabling comparative analyses of protein and nucleic acid composition between organelles isolated from distinct cellular lineages. In animals harbouring a mixture of mutant and wild-type mitochondrial genomes, we use CS-MAP to reveal subtle mosaic patterns of cell-type-specific heteroplasmy across large populations of animals (>10,000 individuals). We demonstrate that the germline is more prone to propagating deleterious mitochondrial genomes than somatic lineages, which we propose is caused by enhanced mtDNA replication in this tissue.
尽管线粒体是普遍存在的细胞器，但它们表现出组织特异性的形态、动态和功能。在这里，我们描述了一种从秀丽隐杆线虫不同组织系统的特定细胞中分离线粒体的稳健方法。细胞特异性线粒体亲和纯化（CS-MAP）可获得完整且功能正常的线粒体，具有极高的纯度和灵敏度（>96%的富集度、>96%的纯度，单细胞和单动物分辨率），从而能够在从不同细胞谱系分离的细胞器之间进行蛋白质和核酸组成的比较分析。在携带混合突变型和野生型线粒体基因组的动物中，我们使用 CS-MAP 揭示了跨越大量动物（>10000 个个体）的细胞类型特异性异质性的微妙镶嵌模式。我们证明生殖细胞比体细胞更容易传播有害的线粒体基因组，我们认为这是由于该组织中 mtDNA 复制增强所致。
