Affinity purification of cell-specific mitochondria from whole animals resolves patterns of genetic mosaicism
Arnaud Ahier, Chuan-Yang Dai, Andrea Tweedie, Ayenachew Bezawork-Geleta, Ina Kirmes, Steven Zuryn
Nature Cell Biology | NATURE PUBLISHING GROUP | Published : 2018
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 s..View full abstract
Awarded by NIH Office of Research Infrastructure Programs
Awarded by NHMRC
Awarded by University of Queensland Early Career Researcher Grant
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.