MITO-Tag Mice enable rapid isolation and multimodal profiling of mitochondria from specific cell types in vivo
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303.full.pdf
Description
Published version
Size
974.7 KB
Format
Adobe PDF
Checksum (MD5)
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Author(s)
Bayraktar, Erol C.
Baudrier, Lou
Özerdem, Ceren
Lewis, Caroline A.
Chan, Sze Ham
Kunchok, Tenzin
Abu-Remaileh, Monther
Cangelosi, Andrew L.
Sabatini, David M.
Birsoy, Kıvanç
Date Issued
December 2018
Journal
Proceedings of the National Academy of Sciences
Publisher
National Academy of Sciences
Citation
Bayraktar, Erol C. et al. "MITO-Tag Mice enable rapid isolation and multimodal profiling of mitochondria from specific cell types in vivo." Proceedings of the National Academy of Sciences 116, 1 (December 2018): 303-312 © 2019 National Academy of Sciences
Version
Final published version
Abstract
Mitochondria are metabolic organelles that are essential for mammalian life, but the dynamics of mitochondrial metabolism within mammalian tissues in vivo remains incompletely understood. While whole-tissue metabolite profiling has been useful for studying metabolism in vivo, such an approach lacks resolution at the cellular and subcellular level. In vivo methods for interrogating organellar metabolites in specific cell types within mammalian tissues have been limited. To address this, we built on prior work in which we exploited a mitochondrially localized 3XHA epitope tag (MITO-Tag) for the fast isolation of mitochondria from cultured cells to generate MITO-Tag Mice. Affording spatiotemporal control over MITO-Tag expression, these transgenic animals enable the rapid, cell-type-specific immunoisolation of mitochondria from tissues, which we verified using a combination of proteomic and metabolomic approaches. Using MITO-Tag Mice and targeted and untargeted metabolite profiling, we identified changes during fasted and refed conditions in a diverse array of mitochondrial metabolites in hepatocytes and found metabolites that behaved differently at the mitochondrial versus whole-tissue level. MITO-Tag Mice should have utility for studying mitochondrial physiology, and our strategy should be generally applicable for studying other mammalian organelles in specific cell types in vivo.
MIT Department
Massachusetts Institute of Technology. Department of Biology
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DOI of Published Version
https://doi.org/10.1073/pnas.1816656115
