Affinity purification of cell-specific mitochondria from whole animals resolves patterns of genetic mosaicism

• Published in: Nature cell biology Vol. 20; no. 3; pp. 352 - 360
• Main Authors: Ahier, Arnaud, Dai, Chuan-Yang, Tweedie, Andrea, Bezawork-Geleta, Ayenachew, Kirmes, Ina, Zuryn, Steven
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 01.03.2018; Nature Publishing Group
• Subjects: 13; 45; 631/136/334/1582/712; 631/1647; 631/208/726/2129; 631/80/642/333; 64; 64/11; 96/63; Affinity; Analysis; Animal genetic engineering; Animals; Biomedical and Life Sciences; Caenorhabditis elegans; Cancer Research; Cell Biology; Developmental Biology; DNA; Genetic engineering; Genomes; Genomics; Heteroplasmy; Life Sciences; Mitochondria; Mitochondrial DNA; Mosaicism; Nucleic acids; Organelles; Protein composition; Purification; Purity; Stem Cells; technical-report
• ISSN: 1465-7392; 1476-4679
• DOI: 10.1038/s41556-017-0023-x

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. Ahier et al. describe a method to isolate intact mitochondria from specific cells in Caenorhabditis elegans and show that the germline is more prone to propagating deleterious mitochondrial genomes than somatic lineages.