A Novel Approach to Measure Mitochondrial Respiration in Frozen Biological Samples

Overview

Journal EMBO J

Specialties Biotechnology
Molecular Biology

Date 2020 May 21

PMID 32432379

Citations 100

Authors

Rebeca Acin-Perez

Ilan Y Benador

Anton Petcherski

Michaela Veliova

Gloria A Benavides

Sylviane Lagarrigue

Arianne Caudal

Laurent Vergnes

Anne N Murphy

Georgios Karamanlidis

Rong Tian

Karen Reue

Jonathan Wanagat

Harold Sacks

Francesca Amati

Victor M Darley-Usmar

Marc Liesa

Ajit S Divakaruni

Linsey Stiles

Orian S Shirihai

Affiliations

Soon will be listed here.

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.

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