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Harmonization of experimental procedures to assess mitochondrial respiration in human permeabilized skeletal muscle fibers.
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.
Afiliação
  • Doerrier C; Oroboros Instruments, Schöpfstrasse 18, 6020, Innsbruck, Austria. Electronic address: carolina.doerrier@gmail.com.
  • Gama-Perez P; Dept Physiological Sciences, Univ Barcelona and Bellvitge Biomedical Research Inst, Spain. Electronic address: p.gamaperez@gmail.com.
  • Pesta D; Inst Clinical Diabetology, German Diabetes Center, Leibniz Center Diabetes Research Heinrich-Heine Univ Düsseldorf, Germany; German Center Diabetes Research, Munich, Neuherberg, Germany; Institute of Aerospace Medicine, German Aerospace Center (DLR), Cologne, Germany; Center for Endocrinology, Diabe
  • Distefano G; Translational Research Institute AdventHealth, Orlando, FL, USA. Electronic address: Giovanna.Distefano@adventhealth.com.
  • Soendergaard SD; Xlab, Dept Biomedical Sciences, Center Healthy Aging, Fac Health Sciences, Denmark. Electronic address: wdz320@alumni.ku.dk.
  • Chroeis KM; Xlab, Dept Biomedical Sciences, Center Healthy Aging, Fac Health Sciences, Denmark. Electronic address: rwn105@alumni.ku.dk.
  • Gonzalez-Franquesa A; The Novo Nordisk Center Basic Metabolic Research, Section Integrative Physiology, Univ Copenhagen, Denmark. Electronic address: agonzalezfranquesa@gmail.com.
  • Goodpaster BH; Translational Research Institute AdventHealth, Orlando, FL, USA. Electronic address: bret.goodpaster@adventhealth.com.
  • Prats C; Dept Biomedical Sciences, Center Healthy Aging, Fac Health Sciences, Denmark; The Core Facility for Integrated Microscopy, Faculty of Health and Medical Sciences, University of Copenhagen, Denmark. Electronic address: cprats@sund.ku.dk.
  • Sales-Pardo M; Dept of Chemical Engineering, Universitat Rovira I Virgili, Tarragona, Spain. Electronic address: marta.sales@urv.cat.
  • Guimera R; Dept of Chemical Engineering, Universitat Rovira I Virgili, Tarragona, Spain; Institució Catalana de Recerca I Estudis Avançats (ICREA), Barcelona, Spain. Electronic address: roger.guimera@urv.cat.
  • Coen PM; Translational Research Institute AdventHealth, Orlando, FL, USA. Electronic address: Paul.Coen@adventhealth.com.
  • Gnaiger E; Oroboros Instruments, Schöpfstrasse 18, 6020, Innsbruck, Austria. Electronic address: erich.gnaiger@oroboros.at.
  • Larsen S; Xlab, Dept Biomedical Sciences, Center Healthy Aging, Fac Health Sciences, Denmark; Clinical Research Centre, Medical University of Bialystok, Bialystok, Poland. Electronic address: stelar@sund.ku.dk.
  • Garcia-Roves PM; Dept Physiological Sciences, Univ Barcelona and Bellvitge Biomedical Research Inst, Spain. Electronic address: pgarciaroves@ub.edu.
Free Radic Biol Med ; 223: 384-397, 2024 Oct.
Article em En | MEDLINE | ID: mdl-39097206
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.
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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Consumo de Oxigênio / Fibras Musculares Esqueléticas / Respiração Celular / Mitocôndrias Musculares Limite: Adult / Humans / Male Idioma: En Revista: Free Radic Biol Med Assunto da revista: BIOQUIMICA / MEDICINA Ano de publicação: 2024 Tipo de documento: Article País de publicação: Estados Unidos
Texto completo
Adicionar na Minha BVS
Imprimir
XML
PubMed Links
Buscar no Google

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Consumo de Oxigênio / Fibras Musculares Esqueléticas / Respiração Celular / Mitocôndrias Musculares Limite: Adult / Humans / Male Idioma: En Revista: Free Radic Biol Med Assunto da revista: BIOQUIMICA / MEDICINA Ano de publicação: 2024 Tipo de documento: Article País de publicação: Estados Unidos