A novel approach to measure mitochondrial respiration in frozen biological samples

被引：154

作者：

Acin-Perez, Rebeca ^{[1
,2
]}

Benador, Ilan Y. ^{[1
,2
,3
]}

Petcherski, Anton ^{[1
,2
]}

Veliova, Michaela ^{[1
,2
,4
]}

Benavides, Gloria A. ^{[5
,6
]}

Lagarrigue, Sylviane ^{[7
]}

Caudal, Arianne ^{[8
]}

Vergnes, Laurent ^{[2
,9
]}

Murphy, Anne N. ^{[10
]}

Karamanlidis, Georgios ^{[11
]}

Tian, Rong ^{[8
]}

Reue, Karen ^{[2
,9
]}

Wanagat, Jonathan ^{[2
,12
]}

Sacks, Harold ^{[1
,2
]}

Amati, Francesca ^{[7
]}

Darley-Usmar, Victor M. ^{[5
,6
]}

Liesa, Marc ^{[1
,2
,4
,13
]}

Divakaruni, Ajit S. ^{[2
,4
]}

Stiles, Linsey ^{[1
,2
]}

Shirihai, Orian S. ^{[1
,2
,3
,4
,13
]}

机构：

[1] Univ Calif Los Angeles, David Geffen Sch Med, Dept Med, Los Angeles, CA 90095 USA

[2] Univ Calif Los Angeles, David Geffen Sch Med, Metab Theme, Los Angeles, CA 90095 USA

[3] Boston Univ, Sch Med, Grad Med Sci, Nutr & Metab, Boston, MA 02118 USA

[4] Univ Calif Los Angeles, Dept Mol & Med Pharmacol, Los Angeles, CA 90095 USA

[5] Univ Alabama Birmingham, Dept Pathol, Birmingham, AL 35294 USA

[6] Univ Alabama Birmingham, Mitochondrial Med Lab, Birmingham, AL USA

[7] Univ Lausanne, Dept Biomed Sci, Lausanne, Switzerland

[8] Univ Washington, Mitochondria & Metab Ctr, Seattle, WA 98195 USA

[9] Univ Calif Los Angeles, David Geffen Sch Med, Dept Human Genet, Los Angeles, CA 90095 USA

[10] Univ Calif San Diego, Dept Pharmacol, San Diego, CA 92103 USA

[11] Amgen Res, Cardiometab Disorders, Thousand Oaks, CA USA

[12] Univ Calif Los Angeles, Dept Med, Div Geriatr, Los Angeles, CA 90024 USA

[13] Univ Calif Los Angeles, Inst Mol Biol, Los Angeles, CA 90024 USA

来源：

EMBO JOURNAL | 2020年 / 39卷 / 13期

基金：

瑞士国家科学基金会;

关键词：

frozen tissue; methodology; mitochondrial content; mitochondrial uncoupled respiration; oxygen consumption; INDUCED LACTIC-ACIDOSIS; OXIDATIVE-PHOSPHORYLATION; MUSCLE; CRYOPRESERVATION; MEMBRANE; PROTEIN; ALAMETHICIN; BIOGENESIS; APOPTOSIS; METFORMIN;

D O I：

10.15252/embj.2019104073

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

070307 [化学生物学]; 071010 [生物化学与分子生物学];

摘要：

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 toIVin one measurement, collected at remote sites or retrospectively in samples residing in tissue biobanks.

引用

页数：18

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