A novel approach to measure mitochondrial respiration in frozen biological samples.

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Ressource 1Download: Front page PostPrint_EMBOJ.pdf (119.47 [Ko])
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Version: Author's accepted manuscript
License: CC BY 4.0
Secondary document(s)
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 R., Benador I.Y., Petcherski A., Veliova M., Benavides G.A., Lagarrigue S., Caudal A., Vergnes L., Murphy A.N., Karamanlidis G., Tian R., Reue K., Wanagat J., Sacks H., Amati F., Darley-Usmar V.M., Liesa M., Divakaruni A.S., Stiles L., Shirihai O.S.
ISSN
1460-2075 (Electronic)
ISSN-L
0261-4189
Publication state
Published
Issued date
01/07/2020
Peer-reviewed
Oui
Volume
39
Number
13
Pages
e104073
Language
english
Notes
Publication types: Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
Publication Status: ppublish
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
Animals, Cryopreservation, Electron Transport Chain Complex Proteins/metabolism, Male, Mice, Mitochondria/metabolism, Oxygen Consumption, Zebrafish/metabolism, Zebrafish Proteins/metabolism, frozen tissue, methodology, mitochondrial content, mitochondrial uncoupled respiration, oxygen consumption
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 9:16
Last modification date
25/04/2021 6:34
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