Technical Report | Published:

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

Nature Cell Biologyvolume 20pages352360 (2018) | Download Citation

Abstract

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.

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Change history

  • 15 February 2018

    In the version of this Technical Report originally published, chromosome representations (indicated by black lines) were missing from Fig. 2a due to a technical error. The corrected version of Fig. 2a is shown below. This has now been amended in all online versions of the Technical Report.

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Acknowledgements

The authors thank R. Tweedale and L. Richards for comments on the manuscript, M. Hilliard and members of the Zuryn laboratory for discussions and comments, M. Hilliard and T. Bredy for sharing reagents and equipment, R. Amor and L. Hammond for support with microscopy, and A. Ho for support with statistics. Some strains were provided by the CGC, which is funded by the NIH Office of Research Infrastructure Programs (P40 OD010440). This work was supported by NHMRC Project Grant 1128381, a University of Queensland Early Career Researcher Grant (UQECR1608181) and a Stafford Fox Senior Research Fellowship to S.Z., and a University of Queensland International Scholarship to C.Y.D.

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Affiliations

  1. Clem Jones Centre for Ageing Dementia Research, Queensland Brain Institute, The University of Queensland, Brisbane, Australia

    • Arnaud Ahier
    • , Chuan-Yang Dai
    • , Andrea Tweedie
    • , Ayenachew Bezawork-Geleta
    • , Ina Kirmes
    •  & Steven Zuryn

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Contributions

A.A. carried out most experiments. C.-Y.D., A.T., A.B.-G., I.K. and S.Z. contributed some experiments. A.A. and S.Z. designed and interpreted experiments and wrote the paper.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Steven Zuryn.

Supplementary information

  1. Supplementary Information

    Supplementary Figures 1–7 and Supplementary References.

  2. Life Sciences Reporting Summary

  3. Supplementary Table 1

    Statistical source data for Figure 6.

  4. Supplementary Table 2

    List of primers used in this study.

  5. Supplementary Table 3

    List of generated transgenic strains used in this study.

  6. Supplementary Table 4

    Antibodies and the working dilutions used in this study.

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DOI

https://doi.org/10.1038/s41556-017-0023-x