A novel approach to measure mitochondrial respiration in frozen biological samples
Open Access
- 20 May 2020
- journal article
- research article
- Published by Springer Science and Business Media LLC in The EMBO Journal
- Vol. 39 (13), e104073
- https://doi.org/10.15252/embj.2019104073
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
Funding Information
- National Institutes of Health (R01 DK099618-05, R01 CA232056-01, R21AG060456-01, R21 AG063373-01, HL110349, U54 DK120342, R56AG060880, R01AG055518, K02AG059847, R21AR072950, R01AA026914-01A1)
- American Diabetes Association (1-19-IBS-049)
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