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Proc Natl Acad Sci U S A. 2019 Jan 2;116(1):303-312. doi: 10.1073/pnas.1816656115. Epub 2018 Dec 12.

MITO-Tag Mice enable rapid isolation and multimodal profiling of mitochondria from specific cell types in vivo.

Author information

1
Laboratory of Metabolic Regulation and Genetics, The Rockefeller University, New York, NY 10065.
2
Whitehead Institute for Biomedical Research, Cambridge, MA 02142.
3
Howard Hughes Medical Institute, Massachusetts Institute of Technology, Cambridge, MA 02139.
4
Department of Biology, Massachusetts Institute of Technology, Cambridge, MA 02139.
5
Koch Institute for Integrative Cancer Research, Cambridge, MA 02139.
6
Broad Institute of Massachusetts Institute of Technology and Harvard, Cambridge, MA 02142.
7
Whitehead Institute for Biomedical Research, Cambridge, MA 02142; sabatini@wi.mit.edu kbirsoy@mail.rockefeller.edu walter.chen@childrens.harvard.edu.
8
Laboratory of Metabolic Regulation and Genetics, The Rockefeller University, New York, NY 10065; sabatini@wi.mit.edu kbirsoy@mail.rockefeller.edu walter.chen@childrens.harvard.edu.

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.

KEYWORDS:

MITO-Tag Mice; lipidomics; metabolomics; mitochondria; proteomics

Comment in

PMID:
30541894
PMCID:
PMC6320505
[Available on 2019-07-02]
DOI:
10.1073/pnas.1816656115
[Indexed for MEDLINE]

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