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A novel approach to measure mitochondrial respiration in frozen biological samples.
Acin-Perez, Rebeca; Benador, Ilan Y; Petcherski, Anton; Veliova, Michaela; Benavides, Gloria A; Lagarrigue, Sylviane; Caudal, Arianne; Vergnes, Laurent; Murphy, Anne N; 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.
Affiliation
  • Acin-Perez R; Department of Medicine, Endocrinology, David Geffen School of Medicine, University of California, Los Angeles, CA, USA.
  • Benador IY; Metabolism Theme, David Geffen School of Medicine, University of California, Los Angeles, CA, USA.
  • Petcherski A; Department of Medicine, Endocrinology, David Geffen School of Medicine, University of California, Los Angeles, CA, USA.
  • Veliova M; Metabolism Theme, David Geffen School of Medicine, University of California, Los Angeles, CA, USA.
  • Benavides GA; Nutrition and Metabolism, Graduate Medical Sciences, Boston University School of Medicine, Boston, MA, USA.
  • Lagarrigue S; Department of Medicine, Endocrinology, David Geffen School of Medicine, University of California, Los Angeles, CA, USA.
  • Caudal A; Metabolism Theme, David Geffen School of Medicine, University of California, Los Angeles, CA, USA.
  • Vergnes L; Department of Medicine, Endocrinology, David Geffen School of Medicine, University of California, Los Angeles, CA, USA.
  • Murphy AN; Metabolism Theme, David Geffen School of Medicine, University of California, Los Angeles, CA, USA.
  • Karamanlidis G; Department of Molecular and Medical Pharmacology, University of California, Los Angeles, CA, USA.
  • Tian R; Department of Pathology and Mitochondrial Medicine Laboratory, University of Alabama at Birmingham, Birmingham, AL, USA.
  • Reue K; Department of Biomedical Sciences, University of Lausanne, Lausanne, Switzerland.
  • Wanagat J; Mitochondria and Metabolism Center, University of Washington, Seattle, WA, USA.
  • Sacks H; Metabolism Theme, David Geffen School of Medicine, University of California, Los Angeles, CA, USA.
  • Amati F; Department of Human Genetics, David Geffen School of Medicine, University of California, Los Angeles, CA, USA.
  • Darley-Usmar VM; Department of Pharmacology, University of California, San Diego, CA, USA.
  • Liesa M; Cardiometabolic Disorders, Amgen Research, Thousand Oaks, CA, USA.
  • Divakaruni AS; Mitochondria and Metabolism Center, University of Washington, Seattle, WA, USA.
  • Stiles L; Metabolism Theme, David Geffen School of Medicine, University of California, Los Angeles, CA, USA.
  • Shirihai OS; Department of Human Genetics, David Geffen School of Medicine, University of California, Los Angeles, CA, USA.
EMBO J ; 39(13): e104073, 2020 07 01.
Article in En | MEDLINE | ID: mdl-32432379
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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Full text: 1 Collection: 01-internacional Database: MEDLINE Main subject: Oxygen Consumption / Zebrafish / Cryopreservation / Zebrafish Proteins / Electron Transport Chain Complex Proteins / Mitochondria Limits: Animals Language: En Journal: EMBO J Year: 2020 Document type: Article Affiliation country:
Fulltext
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Full text: 1 Collection: 01-internacional Database: MEDLINE Main subject: Oxygen Consumption / Zebrafish / Cryopreservation / Zebrafish Proteins / Electron Transport Chain Complex Proteins / Mitochondria Limits: Animals Language: En Journal: EMBO J Year: 2020 Document type: Article Affiliation country: