RRC ID 51549
Author Ahier A, Dai CY, Tweedie A, Bezawork-Geleta A, Kirmes I, Zuryn S.
Title Affinity purification of cell-specific mitochondria from whole animals resolves patterns of genetic mosaicism.
Journal Nat Cell Biol
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.
Volume 20(3)
Pages 352-360
Published 2018-3-1
DOI 10.1038/s41556-017-0023-x
PII 10.1038/s41556-017-0023-x
PMID 29358705
MeSH Animals Animals, Genetically Modified Caenorhabditis elegans / genetics* Caenorhabditis elegans / metabolism Cell Fractionation / methods* Chromatography, Affinity* DNA Replication DNA, Mitochondrial / biosynthesis DNA, Mitochondrial / genetics* Microscopy, Confocal Mitochondria / genetics* Mitochondria / metabolism Mosaicism* Mutation Organ Specificity
IF 20.042
Times Cited 16
Resource
C.elegans tm1779