Doerrier 2024 Free Radic Biol Med
| Doerrier C, Gama-Perez P, Pesta D, Distefano G, Soendergaard SD, Maise Chroeis K, Gonzalez-Franquesa A, Goodpaster BH, Prats C, Sales-Pardo M, Guimera R, Coen PM, Gnaiger E, Larsen S, Garcia-Roves PM (2024) Harmonization of experimental procedures to assess mitochondrial respiration in human permeabilized skeletal muscle fibers. Free Radic Biol Med 223:384-97. https://doi.org/10.1016/j.freeradbiomed.2024.07.039 |
Doerrier Carolina, Gama-Perez Pau, Pesta Dominik, Distefano Giovanna, Soendergaard Stine D, Maise Chroeis Karoline, Gonzalez-Franquesa Alba, Goodpaster Bret H, Prats Clara, Sales-Pardo Marta, Guimera Roger, Coen Paul M, Gnaiger Erich, Larsen Steen, Garcia-Roves Pablo M (2024) Free Radic Biol Med
Abstract: Aim: High-resolution respirometry in human permeabilized muscle fibers is extensively used for analysis of mitochondrial adaptions to nutrition and exercise interventions, and is linked to athletic performance. However, the lack of standardization of experimental conditions limits quantitative inter- and intra-laboratory comparisons.
Methods: In our study, an international team of investigators measured mitochondrial respiration of permeabilized muscle fibers obtained from three biopsies (vastus lateralis) from the same healthy volunteer to avoid inter-individual variability. High-resolution respirometry assays were performed together at the same laboratory to assess whether the heterogenity in published results are due to the effects of respiration media (MiR05 versus Z) with or without the myosin inhibitor blebbistatin at low- and high-oxygen regimes.
Results: Our findings reveal significant differences between respiration media for OXPHOS and ET capacities supported by NADH&succinate-linked substrates at different oxygen concentrations. Respiratory capacities were approximately 1.5-fold higher in MiR05 at high-oxygen regimes compared to medium Z near air saturation. The presence or absence of blebbistatin in human permeabilized muscle fiber preparations was without effect on oxygen flux.
Conclusion: Our study constitutes a basis to harmonize and establish optimum experimental conditions for respirometric studies of permeabilized human skeletal muscle fibers to improve reproducibility. • Keywords: permeabilized skeletal muscle fibers; mitochondrial respiration; high-resolution respirometry; respiration medium MiR05; respiration medium Z; O2 regime; blebbistatin • Bioblast editor: Gnaiger E • O2k-Network Lab: ES Barcelona Garcia-Roves PM, DK Copenhagen Larsen S, US FL Orlando Goodpaster BH, AT Innsbruck Oroboros
Labels: MiParea: Gender, Respiration, Instruments;methods, mt-Structure;fission;fusion, Exercise physiology;nutrition;life style
Organism: Human
Tissue;cell: Skeletal muscle
Preparation: Permeabilized tissue
Regulation: Oxygen kinetics Coupling state: ET, LEAK, OXPHOS Pathway: N, S, NS, ROX HRR: Oxygraph-2k
MiR05
