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Difference between revisions of "Chabi 2019 MitoFit Preprint Arch EA"

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(Created page with "{{MitoFit page name}} {{Publication |title=Chabi BĂ©atrice, Ost M, Gama-Perez P, Dahdah N, Lemieux H, Holody CD, Carpenter RG, Tepp K, Puurand M, Kaambre T, Dubouchaud H, Cort...")
 
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{{Publication
{{Publication
|title=Chabi BĂ©atrice, Ost M, Gama-Perez P, Dahdah N, Lemieux H, Holody CD, Carpenter RG, Tepp K, Puurand M, Kaambre T, Dubouchaud H, Cortade F, Pesta D, Calabria E, Casado M, Fernandez-Ortiz M, Acuña-Castroviejo D, Villena JA, Grefte S, Keijer J, O'Brien K, Sowton A, Murray AJ, Campbell MD, Marcinek DJ, Nollet E, WĂŒst R, Dayanidhi S, Gnaiger E, Doerrier C, Garcia-Roves PM (2019) Generating reference values on mitochondrial respiration in permeabilized muscle fibers. MitoFit Preprint Arch  [[doi:10.26124/mitofit:ea19.MitoEAGLE.0001]].
|title=Chabi BĂ©atrice, Ost M, Gama-Perez P, Dahdah N, Lemieux H, Holody CD, Carpenter RG, Tepp K, Puurand M, Kaambre T, Dubouchaud H, Cortade F, Pesta D, Calabria E, Casado M, Fernandez-Ortiz M, Acuña-Castroviejo D, Villena JA, Grefte S, Keijer J, O'Brien K, Sowton A, Murray AJ, Campbell MD, Marcinek DJ, Nollet E, WĂŒst R, Dayanidhi S, Gnaiger E, Doerrier C, Garcia-Roves PM (2019) Generating reference values on mitochondrial respiration in permeabilized muscle fibers. MitoFit Preprint Arch  [[doi:10.26124/mitofit:ea19.MitoEAGLE.0001]].
|info=[[File:MitoFit Preprint Arch pdf.png|left|160px|link=https://www.mitoeagle.org/images/7/7c/Doerrier_2019_MitoFit_Preprint_Arch_doi_10.26214mitofitea19.MiPSchool.0009.pdf |MitoFit pdf]]  <big><big>'''[https://www.mitoeagle.org/images/7/7c/Doerrier_2019_MitoFit_Preprint_Arch_doi_10.26214mitofitea19.MiPSchool.0009.pdf Inter-laboratory harmonization of respiratory protocols in permeabilized human muscle fibers]'''</big></big>
|info=[[File:MitoFit Preprint Arch pdf.png|left|160px|link=https://www.mitofit.org/images/5/57/Chabi_2019_MitoFit_Preprint_Arch_doi_10.26124mitofitea19.mitoeagle.0001.pdf |MitoFit pdf]]  <big><big>'''[https://www.mitofit.org/images/5/57/Chabi_2019_MitoFit_Preprint_Arch_doi_10.26124mitofitea19.mitoeagle.0001.pdf Generating reference values on mitochondrial respiration in permeabilized muscle fibers]'''</big></big>
|authors=Doerrier C, Gama-Perez P, Distefano G, Pesta D, Soendergaard SD, Chroeis KM, Gonzalez-Franquesa A, Goodpaster BH, Coen P, Larsen S, Gnaiger E, Garcia-Roves PM
|authors=Chabi BĂ©atrice, Ost M, Gama-Perez P, Dahdah N, Lemieux H, Holody CD, Carpenter RG, Tepp K, Puurand M, Kaambre T, Dubouchaud H, Cortade F, Pesta D, Calabria E, Casado M, Fernandez-Ortiz M, Acuña-Castroviejo D, Villena JA, Grefte S, Keijer J, O'Brien K, Sowton A, Murray AJ, Campbell MD, Marcinek DJ, Nollet E, WĂŒst R, Dayanidhi S, Gnaiger E, Doerrier C, Garcia-Roves PM
|year=2019
|year=2019
|journal=MitoFit Preprint Arch
|journal=MitoFit Preprint Arch
|abstract=Version 1 ('''v1''') '''2019-07-04''' [https://www.mitoeagle.org/images/7/7c/Doerrier_2019_MitoFit_Preprint_Arch_doi_10.26214mitofitea19.MiPSchool.0009.pdf doi:10.26124/mitofit:ea19.MiPSchool.0009]
|abstract=Version 1 ('''v1''') '''2019-10-10''' [https://www.mitoeagle.org/images/7/7c/Doerrier_2019_MitoFit_Preprint_Arch_doi_10.26214mitofitea19.MiPSchool.0009.pdf doi:10.26124/mitofit:ea19.MiPSchool.0009]
[[Image:MITOEAGLE-logo.jpg|left|100px|link=http://www.mitoeagle.org/index.php/MitoEAGLE|COST Action MitoEAGLE]]
[[Image:MITOEAGLE-logo.jpg|left|100px|link=http://www.mitoeagle.org/index.php/MitoEAGLE|COST Action MitoEAGLE]]
Permeabilized muscle fibers are extensively used for analysis of mitochondrial function in exercise and pathophysiological studies. Inter- and intra-laboratory comparisons of published results on permeabilized muscle fibers are difficult due to application of different experimental procedures, including sample preparation, substrate-uncoupler-inhibitor titrations (SUIT), respiratory media, and oxygen regimes. Oxygen dependence of mitochondrial respiration in permeabilized fibers (about 100-fold higher p50 compared to small living cells and isolated mitochondria [1]) reveals the requirement of using hyperoxic incubation conditions to avoid oxygen limitation of respiratory capacity [2]. However, controversial results on the oxygen dependence of permeabilized muscle fibers have been reported by different research groups using different respiration media in the presence or absence of the myosin II-specific inhibitor blebbistatin [3,4].  
Permeabilized muscle fibers (pfi) are widely used to assess mitochondrial (mt) respiratory function in skeletal muscle of various models in different physiological and pathological conditions. Facing the numerous data available for mt-respiration from the literature, it remains challenging to determine what the right values are for a specific respiratory protocol. Moreover, mt-respiration values are highly dependent on pfi preparation, which required good technical skills.


In the framework of COST Action MitoEAGLE, our main goals for the current study of permeabilized human muscle fibers are: (1) a comparison of protocols used in different research laboratories, (2) harmonization of results to address the reproducibility crisis [5], (3) evaluation of optimum experimental conditions, and (4) analysis of the causes of experimental variability.
In the frame of COST Action MITOEAGLE, one of the objectives of WG2 is the generation of reference values for mitochondrial respirometry in permeabilized skeletal
muscle sample preparations. The idea is that new researchers in the field follow a reference protocol and check if their values are in an acceptable range. This approach
could serve to test researchers’ technical skills and therefore determine if they are proficient enough to perform their own experiments with confidence. - ''Extended abstract''


We performed a blinded test with human permeabilized skeletal fibers. Six groups from Austria, Denmark, Germany, Spain, and USA measured simultaneously in the same laboratory mitochondrial respiration using high-resolution respirometry (O2k; Oroboros Instruments, Austria) in three human biopsies (vastus lateralis) from the same healthy volunteer sampled on three consecutive days. A total of 96 (32/day) permeabilized fiber preparations were assayed. The wet mass of permeabilized fibers ranged from 0.38 to 2.83 mg per chamber. Protocols were compared at several levels: (1) permeabilized fiber preparation; (2) respiration media MiR05-Kit and Buffer Z in the presence/absence of blebbistatin (25 ”M), covering the most frequently used experimental conditions in the literature; (3) ‘normoxia’ (200-100 ”M) versus hyperoxia (450-250 ”M). The SUIT-008 protocol [6] was applied in all assays. Results were excluded from analysis if the cytochrome c flux control factor, ''FCF<sub>c</sub>'' = (''I''<sub>O2,''c''PM</sub>-''I''<sub>O2,PM</sub>)/''I''<sub>O2,''c''PM</sub>, exceeded 0.1 in the OXPHOS-state (Fig. 1; steps 2D and 2c). For abbreviations see Figure 1 and Gnaiger et al 2019 [7].
== Figures ==
[[File:Chabi 2019 MitoFit Preprint Arch EA Figure 1.png|left|400 px]] '''Figure 1. Substrate-uncoupler-inhibitor titration protocol (SUIT-008 O2 pfi D014).''' Sequential titrations and respiratory states. '''1PM:''' NADH-pathway (N-pathway) in the presence of 5 mM pyruvate and 2 mM malate in the N-LEAK state. '''2D:''' saturating ADP (N-OXPHOS state). '''2c:''' 10 ΌM cytochrome c for evaluating the integrity of the outer mitochondrial membrane. '''3G:''' 10 mM glutamate as an additional NADH-linked substrate (N-OXPHOS state). '''4S:''' 10 mM succinate (NS-OXPHOS capacity). '''5U:''' uncoupler titrations to evaluate the electron transfer- (ET-) capacity (NS-ET capacity). '''6Rot:''' inhibition of CI by rotenone (S-ET capacity). '''7Ama:''' inhibition of CIII by antimycin A (residual oxygen consumption, Rox). Oxygen concentration range in the experiment was maintained between 400-250 ”M O2.


NS-OXPHOS capacity was oxygen-limited under ‘normoxic’ compared to hyperoxic conditions in both media (Figure 2A-D). Blebbistatin did not prevent the decrease of respiration in the ‘normoxic’ regime (Figure 2A and 2C), and exerted minor effects on oxygen flux in both media (Figure 2E-F). These results indicate that oxygen dependence is critical and independent of experimental buffers and blebbistatin (Figure 2A-D). Comparing respiratory capacity in both media under hyperoxic conditions, oxygen flux per mass was higher in MiR05-Kit than in Buffer Z (Figure 2E-F). Evaluation of these trends will be completed based on an in-depth statistical analysis. Our inter-laboratory study provides a basis to harmonize published results on permeabilized human skeletal muscle fibers and establishes guidelines for selecting optimum experimental conditions. - ''Extended abstract''
[[File:Chabi 2019 MitoFit Preprint Arch EA Figure 2.png|left|400 px]] '''Figure 2. NS-OXPHOS capacity of permeabilized soleus muscle fibers.''' NS-OXPHOS capacity was measured in permeabilized soleus muscle fibers from male C57BL/6J mice by 16 research groups. Median with interquartile range show results from individual group with muscle fibers obtained from at least four soleus muscles.
|editor=[[Gnaiger E]] & [[Beno M]] & [[Gnaiger C]]
 
|mipnetlab=AT Innsbruck Oroboros, AT Innsbruck Gnaiger E, US FL Orlando Goodpaster BH, DE Duesseldorf Roden M, DK Copenhagen Dela F, DK Copenhagen Larsen S, ES Barcelona Garcia-Roves PM
== References ==
::::# Lemieux H, Blier PU, Gnaiger E (2017) Remodeling pathway control of mitochondrial respiratory capacity by temperature in mouse heart: electron flow through the Q-junction in permeabilized fibers. Sci Rep 7:2840, DOI:10.1038/s41598-017-02789-8. - https://www.bioblast.at/index.php/SUIT-8_O2_pfi_D14
::::# Baker M (2016) 1,500 scientists lift the lid on reproducibility. Survey sheds light on the ‘crisis’ rocking research. Nature 533:452–4.
 
== Affiliations ==
Chabi BĂ©atrice (1), Ost M (2), Gama-Perez P (3), Dahdah N (3), Lemieux H (4), Holody CD (4), Carpenter RG (4), Tepp K (5), Puurand M (5), Kaambre T (5), Dubouchaud H (6), Cortade F (1), Pesta D (7,8), Calabria E (9), Casado M (1)0, Fernandez-Ortiz M (11), Acuña-Castroviejo D (11), Villena JA (12), Grefte S (13), Keijer J (13), O'Brien K (14), Sowton A (14), Murray AJ (14), Campbell MD (15), Marcinek DJ (15), Nollet E (16), WĂŒst R (16), Dayanidhi S (17), Gnaiger E (18,19), Doerrier C (18), Garcia-Roves PM (3)
::::# DMEM, INRA, Univ Montpellier, France - beatrice.chabi@inra.fr
::::# German Institute of Human Nutrition Potsdam-Rehbruecke, Germany
::::# Dept Physiological Sciences, Univ Barcelona and Bellvitge Biomedical Research Institute (IDIBELL) Spain
::::# Faculty Saint-Jean, Univ Alberta, Canada
::::# Lab of Chemical Biology, National Inst of Chemical Physics and Biophysics, Estonia
::::# Lab Bioénergétique Fondamentale et Appliquée, Univ Grenoble Alpes, INSERM, U1055, France
::::# Institute for Clinical Diabetology, German Diabetes Center, Leibniz Center for Diabetes Research at Heinrich-Heine Univ DĂŒsseldorf, Germany
::::# German Center for Diabetes Research, Munich, Neuherberg, Germany
::::# Dept of Neurological and Movement Sciences, Univ of Verona, Italy
::::# Dept of Molecular and Cellular Pathology and Therapy, Instituto de Biomedicina de Valencia, Spain
::::# Biomedical Research Center, Univ of Granada, Spain
::::# Metabolism and Obesity Lab, Vall d’Hebron Research Inst, Spain
::::# Human and Animal Physiology, Wageningen Univ, The Netherlands;
::::# Dept of Physiology, Development & Neuroscience, Univ of Cambridge, UK
::::# Dept of Radiology, Univ of Washington, South Lake Union, USA
::::# Dept of Human Movement Sciences, Faculty of Behavioural and Movement Sciences, Vrije Univ Amsterdam, The Netherlands
::::# Rehabilitation Institute of Chicago, Feinberg School of Medicine, Northwestern Univ, USA
::::# Oroboros Instruments, Austria
::::# Dept Visceral, Transplant Thoracic Surgery, Daniel Swarovski Research Lab, Medical Univ Innsbruck, Austria
 
== Support ==
{{NextGen-O2k H2020-support}}
[[File:MITOEAGLE-representation.jpg|left|60px|link=http://www.mitoeagle.org/index.php/MitoEAGLE|COST Action MitoEAGLE]]
[[File:COST EU Logo.PNG|left|200px]]
:::: This preprint is based upon work from COST Action CA15203 MitoEAGLE, supported by COST (European Cooperation in Science and Technology).
 
 
|editor=[[Iglesias-Gonzalez J]]
|mipnetlab=
}}
}}
{{Labeling
{{Labeling
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|additional=Preprints,
|additional=Preprints,
}}
}}
== Affiliations ==
Doerrier C (1), Gama-Perez P (2), Distefano G (3), Pesta D (4),(5), Soendergaard SD (6), Chroeis KM (6), Gonzalez-Franquesa A (7), Goodpaster BH (3), Coen P (3), Larsen S (6), Gnaiger E (1),(8), Garcia-Roves PM (2)
::::# Oroboros Instruments, Innsbruck, Austria - [email protected]
::::# Dept Physiological Sciences, Univ Barcelona and Bellvitge Biomedical Research Inst, Spain
::::# Translational Research Inst Metabolism Diabetes, Florida Hospital, Orlando, FL, USA
::::# Inst Clinical Diabetology, German Diabetes Center, Leibniz Center Diabetes Research Heinrich-Heine Univ DĂŒsseldorf
::::# German Center Diabetes Research, Munich, Neuherberg; Germany
::::# Dept Biomedical Sciences, Center Healthy Aging, Fac Health Sciences, Denmark
::::# The Novo Nordisk Center Basic Metabolic Research, Section Integrative Physiology; Univ Copenhagen, Denmark
::::# D Swarovski Research Lab, Dept Visceral, Transplant Thoracic Surgery, Med Univ Innsbruck, Austria
== Support ==
{{NextGen-O2k H2020-support}}
[[File:MITOEAGLE-representation.jpg|left|60px|link=http://www.mitoeagle.org/index.php/MitoEAGLE|COST Action MitoEAGLE]]
[[File:COST EU Logo.PNG|left|200px]]
:::: This preprint is based upon work from COST Action CA15203 MitoEAGLE, supported by COST (European Cooperation in Science and Technology).
== Figures ==
[[File:Doerrier_Figure1.jpg|left|400 px]] '''Figure 1. Substrate-uncoupler-inhibitor titration protocol (SUIT-008 O2 pfi D014).''''''Bold text''' Sequential titrations and respiratory states. '''1PM''': NADH-pathway (N-pathway) in the presence of 5 mM pyruvate and 2 mM malate in the N-LEAK state. '''2D''''''Bold text''': saturating ADP (N-OXPHOS state). '''2c''': 10 ”M cytochrome c for evaluating the integrity of the outer mitochondrial membrane. '''3G''': 10 mM glutamate as an additional NADH-linked substrate (N-OXPHOS state). '''4S''': 10 mM succinate (NS-OXPHOS capacity). '''5U''': uncoupler titrations to evaluate the electron transfer- (ET-) capacity (NS-ET capacity). '''6Rot''': inhibition of CI by rotenone (S-ET capacity). '''7Ama''': inhibition of CIII by antimycin A (residual oxygen consumption, ''Rox'').
[[File:Doerrier_Figure2.jpg|left|400 px]] '''Figure 2. The effect of oxygen concentration and blebbistatin on mitochondrial respiration of permeabilized human skeletal muscle fibers in MiR05-Kit (A, B) and Buffer Z (C, D).''' Mass-specific NS-OXPHOS capacity (based on wet mass) supported by pyruvate, malate, glutamate and succinate. ('''E, F''') Comparison of the two media at hyperoxia in the presence and absence of blebbistatin. A biopsy was taken on three consecutive days from the same person. Scatter plots and median with interquartile range show results from individual chambers (''n'' = 8 to 10) with muscle fibers obtained from the three biopsies.
== References ==
::::# Scandurra FM, Gnaiger E (2010) Cell respiration under hypoxia: facts and artefacts in mitochondrial oxygen kinetics. Adv Exp Med Biol 662:7-25.
::::# Gnaiger E (2003) Oxygen conformance of cellular respiration. A perspective of mitochondrial physiology. Adv Exp Med Biol 543:39-55.
::::# Perry CG, Kane DA, Lin CT, Kozy R, Cathey BL, Lark DS, Kane CL, Brophy PM, Gavin TP, Anderson EJ, Neufer PD (2011) Inhibiting myosin-ATPase reveals a dynamic range of mitochondrial respiratory control in skeletal muscle. Biochem J 437:215-22.
::::# Bezuidenhout N, Doerrier C, Droescher S, Ojuka E, Gnaiger E (2016) Comparison of oxygen dependence of respiration in permeabilized mouse skeletal muscle fibers in two respiration media, MiR06Cr and Buffer Z containing Ctl, Cr and Blebbistatin. Abstract MitoFit Science Camp 2016.
::::# Baker M (2016) 1,500 scientists lift the lid on reproducibility. Survey sheds light on the ‘crisis’ rocking research. Nature 533:452–4.
::::# [[SUIT-008_O2_pfi_D014]]
::::# Gnaiger E, Aasander Frostner E, Abdul Karim N, Abumrad NA, Acuna-Castroviejo D, Adiele RC, et al (2019) Mitochondrial respiratory states and rates. MitoFit Preprint Arch doi:10.26124/mitofit:190001.v4.

Revision as of 14:30, 10 October 2019


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Chabi 2019 MitoFit Preprint Arch EA

Publications in the MiPMap
Chabi BĂ©atrice, Ost M, Gama-Perez P, Dahdah N, Lemieux H, Holody CD, Carpenter RG, Tepp K, Puurand M, Kaambre T, Dubouchaud H, Cortade F, Pesta D, Calabria E, Casado M, Fernandez-Ortiz M, Acuña-Castroviejo D, Villena JA, Grefte S, Keijer J, O'Brien K, Sowton A, Murray AJ, Campbell MD, Marcinek DJ, Nollet E, WĂŒst R, Dayanidhi S, Gnaiger E, Doerrier C, Garcia-Roves PM (2019) Generating reference values on mitochondrial respiration in permeabilized muscle fibers. MitoFit Preprint Arch doi:10.26124/mitofit:ea19.MitoEAGLE.0001.

»

MitoFit pdf

Generating reference values on mitochondrial respiration in permeabilized muscle fibers

Chabi BĂ©atrice, Ost M, Gama-Perez P, Dahdah N, Lemieux H, Holody CD, Carpenter RG, Tepp K, Puurand M, Kaambre T, Dubouchaud H, Cortade F, Pesta D, Calabria E, Casado M, Fernandez-Ortiz M, Acuña-Castroviejo D, Villena JA, Grefte S, Keijer J, O'Brien K, Sowton A, Murray AJ, Campbell MD, Marcinek DJ, Nollet E, WĂŒst R, Dayanidhi S, Gnaiger E, Doerrier C, Garcia-Roves PM (2019) MitoFit Preprint Arch

Abstract: Version 1 (v1) 2019-10-10 doi:10.26124/mitofit:ea19.MiPSchool.0009

COST Action MitoEAGLE

Permeabilized muscle fibers (pfi) are widely used to assess mitochondrial (mt) respiratory function in skeletal muscle of various models in different physiological and pathological conditions. Facing the numerous data available for mt-respiration from the literature, it remains challenging to determine what the right values are for a specific respiratory protocol. Moreover, mt-respiration values are highly dependent on pfi preparation, which required good technical skills.

In the frame of COST Action MITOEAGLE, one of the objectives of WG2 is the generation of reference values for mitochondrial respirometry in permeabilized skeletal muscle sample preparations. The idea is that new researchers in the field follow a reference protocol and check if their values are in an acceptable range. This approach could serve to test researchers’ technical skills and therefore determine if they are proficient enough to perform their own experiments with confidence. - Extended abstract

Figures

Chabi 2019 MitoFit Preprint Arch EA Figure 1.png

Figure 1. Substrate-uncoupler-inhibitor titration protocol (SUIT-008 O2 pfi D014). Sequential titrations and respiratory states. 1PM: NADH-pathway (N-pathway) in the presence of 5 mM pyruvate and 2 mM malate in the N-LEAK state. 2D: saturating ADP (N-OXPHOS state). 2c: 10 ΌM cytochrome c for evaluating the integrity of the outer mitochondrial membrane. 3G: 10 mM glutamate as an additional NADH-linked substrate (N-OXPHOS state). 4S: 10 mM succinate (NS-OXPHOS capacity). 5U: uncoupler titrations to evaluate the electron transfer- (ET-) capacity (NS-ET capacity). 6Rot: inhibition of CI by rotenone (S-ET capacity). 7Ama: inhibition of CIII by antimycin A (residual oxygen consumption, Rox). Oxygen concentration range in the experiment was maintained between 400-250 ”M O2.

Chabi 2019 MitoFit Preprint Arch EA Figure 2.png

Figure 2. NS-OXPHOS capacity of permeabilized soleus muscle fibers. NS-OXPHOS capacity was measured in permeabilized soleus muscle fibers from male C57BL/6J mice by 16 research groups. Median with interquartile range show results from individual group with muscle fibers obtained from at least four soleus muscles.

References

  1. Lemieux H, Blier PU, Gnaiger E (2017) Remodeling pathway control of mitochondrial respiratory capacity by temperature in mouse heart: electron flow through the Q-junction in permeabilized fibers. Sci Rep 7:2840, DOI:10.1038/s41598-017-02789-8. - https://www.bioblast.at/index.php/SUIT-8_O2_pfi_D14
  2. Baker M (2016) 1,500 scientists lift the lid on reproducibility. Survey sheds light on the ‘crisis’ rocking research. Nature 533:452–4.

Affiliations

Chabi BĂ©atrice (1), Ost M (2), Gama-Perez P (3), Dahdah N (3), Lemieux H (4), Holody CD (4), Carpenter RG (4), Tepp K (5), Puurand M (5), Kaambre T (5), Dubouchaud H (6), Cortade F (1), Pesta D (7,8), Calabria E (9), Casado M (1)0, Fernandez-Ortiz M (11), Acuña-Castroviejo D (11), Villena JA (12), Grefte S (13), Keijer J (13), O'Brien K (14), Sowton A (14), Murray AJ (14), Campbell MD (15), Marcinek DJ (15), Nollet E (16), WĂŒst R (16), Dayanidhi S (17), Gnaiger E (18,19), Doerrier C (18), Garcia-Roves PM (3)

  1. DMEM, INRA, Univ Montpellier, France - [email protected]
  2. German Institute of Human Nutrition Potsdam-Rehbruecke, Germany
  3. Dept Physiological Sciences, Univ Barcelona and Bellvitge Biomedical Research Institute (IDIBELL) Spain
  4. Faculty Saint-Jean, Univ Alberta, Canada
  5. Lab of Chemical Biology, National Inst of Chemical Physics and Biophysics, Estonia
  6. Lab Bioénergétique Fondamentale et Appliquée, Univ Grenoble Alpes, INSERM, U1055, France
  7. Institute for Clinical Diabetology, German Diabetes Center, Leibniz Center for Diabetes Research at Heinrich-Heine Univ DĂŒsseldorf, Germany
  8. German Center for Diabetes Research, Munich, Neuherberg, Germany
  9. Dept of Neurological and Movement Sciences, Univ of Verona, Italy
  10. Dept of Molecular and Cellular Pathology and Therapy, Instituto de Biomedicina de Valencia, Spain
  11. Biomedical Research Center, Univ of Granada, Spain
  12. Metabolism and Obesity Lab, Vall d’Hebron Research Inst, Spain
  13. Human and Animal Physiology, Wageningen Univ, The Netherlands;
  14. Dept of Physiology, Development & Neuroscience, Univ of Cambridge, UK
  15. Dept of Radiology, Univ of Washington, South Lake Union, USA
  16. Dept of Human Movement Sciences, Faculty of Behavioural and Movement Sciences, Vrije Univ Amsterdam, The Netherlands
  17. Rehabilitation Institute of Chicago, Feinberg School of Medicine, Northwestern Univ, USA
  18. Oroboros Instruments, Austria
  19. Dept Visceral, Transplant Thoracic Surgery, Daniel Swarovski Research Lab, Medical Univ Innsbruck, Austria

Support

Template NextGen-O2k.jpg
COST Action MitoEAGLE
COST EU Logo.PNG
This preprint is based upon work from COST Action CA15203 MitoEAGLE, supported by COST (European Cooperation in Science and Technology).

‱ Bioblast editor: Iglesias-Gonzalez J


Labels: MiParea: Respiration, Instruments;methods 


Organism: Human  Tissue;cell: Skeletal muscle  Preparation: Permeabilized tissue 


Coupling state: LEAK, OXPHOS, ET  Pathway: N, S, NS, ROX  HRR: Oxygraph-2k 

Preprints