A novel approach to measure mitochondrial respiration in frozen biological samples

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License: CC BY 4.0
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Download: Acin_Perez_EMBOjournal_2020.pdf (2547.43 [Ko])
State: Public
Version: Final published version
License: CC BY 4.0
Serval ID
serval:BIB_A152608B8CD2
Type
Article: article from journal or magazin.
Collection
Publications
Institution
Title
A novel approach to measure mitochondrial respiration in frozen biological samples
Journal
The EMBO Journal
Author(s)
Acin-Perez Rebeca, Benador Ilan Y, Petcherski Anton, Veliova Michaela, Benavides Gloria A, Lagarrigue Sylviane, Caudal Arianne, Vergnes Laurent, Murphy Anne M, Karamanlidis Georgios, Tian Rong, Reue Karen, Wanagat Jonathan, Sacks Harold, Amati Francesca, Darley-Usmar Victor M, Liesa Marc, Divakaruni Ajit S, Stiles Linsey, Shirihai Orian S
ISSN
0261-4189
1460-2075
Publication state
Published
Issued date
20/05/2020
Peer-reviewed
Oui
Language
english
Abstract
Respirometry is the gold standard measurement of mitochondrial oxidative function, as it reflects the activity of the electron transport chain complexes working together. However, the requirement for freshly isolated mitochondria hinders the feasibility of respirometry in multi-site clinical studies and retrospective studies. Here, we describe a novel respirometry approach suited for frozen samples by restoring electron transfer components lost during freeze/thaw and correcting for variable permeabilization of mitochondrial membranes. This approach preserves 90-95% of the maximal respiratory capacity in frozen samples and can be applied to isolated mitochondria, permeabilized cells, and tissue homogenates with high sensitivity. We find that primary changes in mitochondrial function, detected in fresh tissue, are preserved in frozen samples years after collection. This approach will enable analysis of the integrated function of mitochondrial Complexes I to IV in one measurement, collected at remote sites or retrospectively in samples residing in tissue biobanks.
Keywords
General Biochemistry, Genetics and Molecular Biology, General Immunology and Microbiology, General Neuroscience, Molecular Biology
Pubmed
Web of science
Open Access
Yes
Funding(s)
Swiss National Science Foundation / Projects / 320030_170062
Swiss National Science Foundation / Projects / 310030-188789
Create date
02/06/2020 8:16
Last modification date
11/06/2020 5:21
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