Author, as appears in the article.: Doerrier, Carolina; Gama-Perez, Pau; Pesta, Dominik; Distefano, Giovanna; Soendergaard, Stine D; Chroeis, Karoline Maise; Gonzalez-Franquesa, Alba; Goodpaster, Bret H; Prats, Clara; Sales-Pardo, Marta; Guimera, Roger; Coen, Paul M; Gnaiger, Erich; Larsen, Steen; Garcia-Roves, Pablo M
Department: Enginyeria Química
URV's Author/s: Guimera Manrique, Roger / Sales Pardo, Marta
Keywords: Small biopsies Responses Respiration medium z Respiration medium mir05 Phosphorylation Permeabilized skeletal muscle fibers Performance Oxygen consumption Oxygen Oxphos protocols Oxidative phosphorylation O2 regim O(2) regime O regime 2 Muscle fibers, skeletal Mitochondrial respiration Mitochondria, muscle Male Humans Human-cells High-resolution respirometry Heterocyclic compounds, 4 or more rings Healthy volunteers H2o2 emission Exercise Cell respiration Capacit Blebbistatin Adult Adaptations
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 ETcapacities 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.
Thematic Areas: Zootecnia / recursos pesqueiros Química Psicología Physiology (medical) Odontología Nutrição Medicina veterinaria Medicina iii Medicina ii Medicina i Materiais Interdisciplinar General medicine Farmacia Engenharias iv Enfermagem Endocrinology & metabolism Educação física Ciências biológicas iii Ciências biológicas ii Ciências biológicas i Ciências ambientais Ciência de alimentos Biotecnología Biodiversidade Biochemistry & molecular biology Biochemistry Astronomia / física
licence for use: https://creativecommons.org/licenses/by/3.0/es/
Author's mail: roger.guimera@urv.cat marta.sales@urv.cat
Author identifier: 0000-0002-3597-4310 0000-0002-8140-6525
Record's date: 2024-10-19
Papper version: info:eu-repo/semantics/publishedVersion
Link to the original source: https://www.sciencedirect.com/science/article/pii/S0891584924005859?dgcid=rss_sd_all
Licence document URL: https://repositori.urv.cat/ca/proteccio-de-dades/
Papper original source: Free Radical Biology And Medicine. 223 384-397
APA: Doerrier, Carolina; Gama-Perez, Pau; Pesta, Dominik; Distefano, Giovanna; Soendergaard, Stine D; Chroeis, Karoline Maise; Gonzalez-Franquesa, Alba; Go (2024). Harmonization of experimental procedures to assess mitochondrial respiration in human permeabilized skeletal muscle fibers. Free Radical Biology And Medicine, 223(), 384-397. DOI: 10.1016/j.freeradbiomed.2024.07.039
Article's DOI: 10.1016/j.freeradbiomed.2024.07.039
Entity: Universitat Rovira i Virgili
Journal publication year: 2024
Publication Type: Journal Publications
Repositori URV