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

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

Journal

The EMBO journal
ISSN: 1460-2075
Titre abrégé: EMBO J
Pays: England
ID NLM: 8208664

Informations de publication

Date de publication:
01 07 2020
Historique:
received: 21 11 2019
revised: 11 03 2020
accepted: 19 03 2020
pubmed: 21 5 2020
medline: 7 4 2021
entrez: 21 5 2020
Statut: ppublish

Résumé

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.

Identifiants

DOI: 10.15252/embj.2019104073 PMID: 32432379 PMC: PMC7327496
pubmed: 32432379
doi: 10.15252/embj.2019104073
pmc: PMC7327496
doi:

Substances chimiques

Electron Transport Chain Complex Proteins 0
Zebrafish Proteins 0

Types de publication

Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't

Langues

eng

Sous-ensembles de citation

IM

Pagination

e104073

Subventions

Organisme : NHLBI NIH HHS
ID : R01 HL142628
Pays : United States
Organisme : NCI NIH HHS
ID : R01 CA232056
Pays : United States
Organisme : NHLBI NIH HHS
ID : R01 HL129510
Pays : United States
Organisme : Swiss National Science Foundation
ID : 320030_170062
Pays : Switzerland
Organisme : NIAMS NIH HHS
ID : R21 AR072950
Pays : United States
Organisme : University Alabama Birmingham
ID : P30 G050886
Organisme : HHS | National Institutes of Health (NIH)
ID : R01AG055518
Organisme : NIA NIH HHS
ID : K02 AG059847
Pays : United States
Organisme : HHS | National Institutes of Health (NIH)
ID : R01AA026914-01A1
Organisme : NIA NIH HHS
ID : R21 AG063373
Pays : United States
Organisme : HHS | National Institutes of Health (NIH)
ID : R56AG060880
Organisme : HHS | National Institutes of Health (NIH)
ID : R01 DK099618-05
Organisme : NIDDK NIH HHS
ID : U54 DK120342
Pays : United States
Organisme : NIA NIH HHS
ID : R01 AG055518
Pays : United States
Organisme : NIA NIH HHS
ID : R56 AG060880
Pays : United States
Organisme : NIDDK NIH HHS
ID : R01 DK099618
Pays : United States
Organisme : NHLBI NIH HHS
ID : R01 HL110349
Pays : United States
Organisme : NIA NIH HHS
ID : P30 AG028748
Pays : United States
Organisme : NIDDK NIH HHS
ID : P30 DK079337
Pays : United States
Organisme : NIAAA NIH HHS
ID : R01 AA026914
Pays : United States
Organisme : American Diabetes Association (ADA)
ID : 1-19-IBS-049
Organisme : HHS | National Institutes of Health (NIH)
ID : HL110349
Organisme : HHS | National Institutes of Health (NIH)
ID : R01 CA232056-01
Organisme : NIA NIH HHS
ID : R21 AG060456
Pays : United States

Informations de copyright

© 2020 The Authors. Published under the terms of the CC BY 4.0 license.

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Auteurs

Rebeca Acin-Perez (R)

Department of Medicine, Endocrinology, David Geffen School of Medicine, University of California, Los Angeles, CA, USA.
Metabolism Theme, David Geffen School of Medicine, University of California, Los Angeles, CA, USA.
ORCID: 0000-0001-9553-8337

Ilan Y Benador (IY)

Department of Medicine, Endocrinology, David Geffen School of Medicine, University of California, Los Angeles, CA, USA.
Metabolism Theme, David Geffen School of Medicine, University of California, Los Angeles, CA, USA.
Nutrition and Metabolism, Graduate Medical Sciences, Boston University School of Medicine, Boston, MA, USA.

Anton Petcherski (A)

Department of Medicine, Endocrinology, David Geffen School of Medicine, University of California, Los Angeles, CA, USA.
Metabolism Theme, David Geffen School of Medicine, University of California, Los Angeles, CA, USA.

Michaela Veliova (M)

Department of Medicine, Endocrinology, David Geffen School of Medicine, University of California, Los Angeles, CA, USA.
Metabolism Theme, David Geffen School of Medicine, University of California, Los Angeles, CA, USA.
Department of Molecular and Medical Pharmacology, University of California, Los Angeles, CA, USA.

Gloria A Benavides (GA)

Department of Pathology and Mitochondrial Medicine Laboratory, University of Alabama at Birmingham, Birmingham, AL, USA.

Sylviane Lagarrigue (S)

Department of Biomedical Sciences, University of Lausanne, Lausanne, Switzerland.

Arianne Caudal (A)

Mitochondria and Metabolism Center, University of Washington, Seattle, WA, USA.

Laurent Vergnes (L)

Metabolism Theme, David Geffen School of Medicine, University of California, Los Angeles, CA, USA.
Department of Human Genetics, David Geffen School of Medicine, University of California, Los Angeles, CA, USA.

Anne N Murphy (AN)

Department of Pharmacology, University of California, San Diego, CA, USA.
ORCID: 0000-0002-5222-9902

Georgios Karamanlidis (G)

Cardiometabolic Disorders, Amgen Research, Thousand Oaks, CA, USA.

Rong Tian (R)

Mitochondria and Metabolism Center, University of Washington, Seattle, WA, USA.

Karen Reue (K)

Metabolism Theme, David Geffen School of Medicine, University of California, Los Angeles, CA, USA.
Department of Human Genetics, David Geffen School of Medicine, University of California, Los Angeles, CA, USA.

Jonathan Wanagat (J)

Metabolism Theme, David Geffen School of Medicine, University of California, Los Angeles, CA, USA.
Department of Medicine, Division of Geriatrics, University of California, Los Angeles, CA, USA.

Harold Sacks (H)

Department of Medicine, Endocrinology, David Geffen School of Medicine, University of California, Los Angeles, CA, USA.
Metabolism Theme, David Geffen School of Medicine, University of California, Los Angeles, CA, USA.

Francesca Amati (F)

Department of Biomedical Sciences, University of Lausanne, Lausanne, Switzerland.

Victor M Darley-Usmar (VM)

Department of Pathology and Mitochondrial Medicine Laboratory, University of Alabama at Birmingham, Birmingham, AL, USA.

Marc Liesa (M)

Department of Medicine, Endocrinology, David Geffen School of Medicine, University of California, Los Angeles, CA, USA.
Metabolism Theme, David Geffen School of Medicine, University of California, Los Angeles, CA, USA.
Department of Molecular and Medical Pharmacology, University of California, Los Angeles, CA, USA.
Molecular Biology Institute, UCLA, Los Angeles, CA, USA.
ORCID: 0000-0002-5909-8570

Ajit S Divakaruni (AS)

Metabolism Theme, David Geffen School of Medicine, University of California, Los Angeles, CA, USA.
Department of Molecular and Medical Pharmacology, University of California, Los Angeles, CA, USA.
ORCID: 0000-0002-2528-9651

Linsey Stiles (L)

Department of Medicine, Endocrinology, David Geffen School of Medicine, University of California, Los Angeles, CA, USA.
Metabolism Theme, David Geffen School of Medicine, University of California, Los Angeles, CA, USA.
ORCID: 0000-0002-1514-458X

Orian S Shirihai (OS)

Department of Medicine, Endocrinology, David Geffen School of Medicine, University of California, Los Angeles, CA, USA.
Metabolism Theme, David Geffen School of Medicine, University of California, Los Angeles, CA, USA.
Nutrition and Metabolism, Graduate Medical Sciences, Boston University School of Medicine, Boston, MA, USA.
Department of Molecular and Medical Pharmacology, University of California, Los Angeles, CA, USA.
Molecular Biology Institute, UCLA, Los Angeles, CA, USA.
ORCID: 0000-0001-8466-3431

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Classifications MeSH

questionsmedicales.fr › Eucaryotes › Animaux › A novel approach to measure mitochondrial...
questionsmedicales.fr › Techniques d'investigation › Conservation biologique › Cryoconservation › A novel approach to measure mitochondrial...
questionsmedicales.fr › Acides aminés, peptides et protéines › Protéines › Protéines membranaires › Complexe enzymatique de la chaine respiratoire mitochondriale › A novel approach to measure mitochondrial...
questionsmedicales.fr › Eucaryotes › Animaux › Chordés › Vertébrés › Mammifères › Eutheria › Rodentia › Muridae › Murinae › Souris › A novel approach to measure mitochondrial...
questionsmedicales.fr › Cellules › Structures cellulaires › Fractions subcellulaires › Mitochondries › A novel approach to measure mitochondrial...
questionsmedicales.fr › Métabolisme › Consommation d'oxygène › A novel approach to measure mitochondrial...
questionsmedicales.fr › Eucaryotes › Animaux › Chordés › Vertébrés › Poissons › Cypriniformes › Cyprinidae › Danio zébré › A novel approach to measure mitochondrial...
questionsmedicales.fr › Acides aminés, peptides et protéines › Protéines › Protéines de poisson › Protéines de poisson-zèbre › A novel approach to measure mitochondrial...