IOC recommended reading

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IOC recommended reading

COST Action MitoEAGLE

MitoEAGLE preprint

  • 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. - »MitoFit Preprint Arch«


O2k-Workshops.png
O2k-Publications in the MiPMap


Workshop material

  • Doerrier C, Garcia-Souza LF, Krumschnabel G, Wohlfarter Y, Mészáros AT, Gnaiger E (2018) High-Resolution FluoRespirometry and OXPHOS protocols for human cells, permeabilized fibres from small biopsies of muscle, and isolated mitochondria. Methods Mol Biol 1782 (Palmeira CM, Moreno AJ, eds): Mitochondrial Bioenergetics, 978-1-4939-7830-4. - »Bioblast link«
  • Gnaiger E (2014) Mitochondrial pathways and respiratory control. An introduction to OXPHOS analysis. 4th ed. Mitochondr Physiol Network 19.12. Oroboros MiPNet Publications, Innsbruck:80 pp. - »Bioblast link« - A new edition is in the pipeline, with updated terminology!
  • Makrecka-Kuka M, Krumschnabel G, Gnaiger E (2015) High-resolution respirometry for simultaneous measurement of oxygen and hydrogen peroxide fluxes in permeabilized cells, tissue homogenate and isolated mitochondria. Biomolecules 5:1319-38. »Bioblast pdf«
  • Krumschnabel G, Eigentler A, Fasching M, Gnaiger E (2014) Use of safranin for the assessment of mitochondrial membrane potential by high-resolution respirometry and fluorometry. Methods Enzymol 542:163-81. - »Bioblast link«
  • O2k-Manual.pdf: O2k-Manual for DatLab 7.


HRR: high-resolution respirometry and SUIT protocols

  • Doerrier C, Garcia-Souza LF, Krumschnabel G, Wohlfarter Y, Mészáros AT, Gnaiger E (2018) High-Resolution FluoRespirometry and OXPHOS protocols for human cells, permeabilized fibres from small biopsies of muscle, and isolated mitochondria. Methods Mol Biol 1782 (Palmeira CM, Moreno AJ, eds): Mitochondrial Bioenergetics, 978-1-4939-7830-4. - »Bioblast link«
  • 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. - »Bioblast link«
» Updated terminology with SUIT protocols.
» NADH-linked or N-pathways (CI-entry into Q); succinate-linked or S-pathway (CII-entry into Q); NS-pathway (convergent CI&II-entry into Q).
  • Gnaiger E (2014) Mitochondrial pathways and respiratory control. An introduction to OXPHOS analysis. 4th ed. Mitochondr Physiol Network 19.12. Oroboros MiPNet Publications, Innsbruck:80 pp. - »Bioblast link«
» A hard copy of this monograph on mitochondrial physiology will be included in the workshop material. The Blue Book is available on-line free of charge and provides a background on OXPHOS analysis:
» From oxygen concentration to oxygen flux •• Membrane potential: electric and chemical forces •• Coupling and efficiency •• Biochemical thresholds
» Respiratory states
» MitoPathways - the electron transfer-pathway
» Normalization of mitochondrial respiration: coupling control and pathway control.


  • Gnaiger E (2008) Polarographic oxygen sensors, the oxygraph and high-resolution respirometry to assess mitochondrial function. In: Mitochondrial Dysfunction in Drug-Induced Toxicity (Dykens JA, Will Y, eds) John Wiley:327-52. - »Bioblast link«
» Methodological introduction, comparison of HRR with traditional oxygraphy.
» Dramatically different: high-resolution respirometry - the Oroboros O2k.
» Calibration of polarographic oxygen sensors and oxygen concentration in respiration media at air saturation.
» From Oxygraph slopes to respiratory flux corrected for background effects.
» Phosphorylation or coupling control protocol (PCP, CCP) with intact cells.
» Titration steps of the CCP.
» Experimental example.
» Flux control ratios from the CCP.
» Intact cells, permeabilized cells and tissue, or isolated mitochondria?




OXPHOS ROUTINE ETS LEAK ROX


O2k-FluoRespirometer

O2k-FluoRespirometry

  • Makrecka-Kuka M, Krumschnabel G, Gnaiger E (2015) High-resolution respirometry for simultaneous measurement of oxygen and hydrogen peroxide fluxes in permeabilized cells, tissue homogenate and isolated mitochondria. Biomolecules 5:1319-38. »Bioblast pdf«
  • Krumschnabel G, Eigentler A, Fasching M, Gnaiger E (2014) Use of safranin for the assessment of mitochondrial membrane potential by high-resolution respirometry and fluorometry. Methods Enzymol 542:163-81. - »Bioblast link«
  • Krumschnabel G, Fontana-Ayoub M, Sumbalova Z, Heidler J, Gauper K, Fasching M, Gnaiger E (2015) Simultaneous high-resolution measurement of mitochondrial respiration and hydrogen peroxide production. Methods Mol Biol 1264:245-61. - »Bioblast link«
  • Eigentler A, Fontana-Ayoub M, Gnaiger E (2013) O2k-Fluorometry: HRR and H2O2 production in mouse cardiac tissue homogenate. Mitochondr Physiol Network 18.05(01):1-6. - »Bioblast link«


TPP/ISE

  • O2k-MultiSensor System with ion selective electrodes (ISE) MiPNet15.03
  • Determination of membrane potential using TPP+ and an ion selective electrode (ISE) MiPNet14.05
  • Rottenberg H (1984) Membrane potential and surface potential in mitochondria: uptake and binding of lipophilic cations. J Membr Biol 81:127-38.


Publications in the MiPMap

Further reading

  • Gnaiger E (2009) Capacity of oxidative phosphorylation in human skeletal muscle. New perspectives of mitochondrial physiology. Int J Biochem Cell Biol 41: 1837-1845. - »Bioblast link«
» Respirometry with permeabilized fibres and isolated mitochondria.
» Convergent NS (CI&II) electron input and OXPHOS capacity.
» Tissue-OXPHOS capacity in human permeabilized muscle fibres and isolated mitochondria.
» Tissue-OXPHOS capacity and functional diversity.


  • Pesta D, Gnaiger E (2012) High-resolution respirometry. OXPHOS protocols for human cells and permeabilized fibres from small biopisies of human muscle. Methods Mol Biol 810: 25-58. - »Bioblast link«
» Introduction to high-resolution respirometry (HRR), SUIT protocols and flux control ratios.


  • Scandurra FM, Gnaiger E (2010) Cell respiration under hypoxia: Facts and artefacts in mitochondrial oxygen kinetics. Adv Exp Med Biol 662: 7-25. - »Bioblast link«
» Isolated mitochondria and intact cells.
» Nanomolar resolution of oxygen concentration.
» Effects of cell density and activity.
» Closed versus open systems.
» p50 in vivo.


  • Gnaiger E, Kuznetsov AV, Schneeberger S, Seiler R, Brandacher G, Steurer W, Margreiter R (2000) Mitochondria in the cold. In: Life in the Cold (Heldmaier G, Klingenspor M, eds) Springer, Heidelberg, Berlin, New York:431-42. - »Bioblast link«
» Isolated mitochondria and permeabilized muscle fibers.
» Optimization of mitochondrial cold storage • Mitochondrial respiration medium, MiR05 • Mitochondrial cold ischemia-reperfusion injury.


  • Renner K, Amberger A, Konwalinka G, Kofler R, Gnaiger E (2003) Changes of mitochondrial respiration, mitochondrial content and cell size after induction of apoptosis in leukemia cells. Biochim Biophys Acta 1642:115-23. - »Bioblast link«
» Intact cells.
» Cytochrome c oxidase.
» Cytochrome c test.
» Respiration per million cells, per citrate synthase, per mg protein, or per cytochrome c oxidase activity.


  • Gnaiger E (2001) Bioenergetics at low oxygen: dependence of respiration and phosphorylation on oxygen and adenosine diphosphate supply. Respir Physiol 128:277-97. - »Bioblast link«
» A detailed introduction into high-resolution respirometry with particular emphasis on kinetics and measurements at low oxygen
» Mitochondrial kinetics measured by high-resolution respirometry
» Calibrations and corrections for response time and instrumental background
» Steady-state injection respirometry
» Mitochondrial respiratory control at low oxygen
» Apparent oxygen affinity and catalytic efficiency of mitochondrial respiration
» Effect of ADP and oxygen limitation on ADP/O2 flux ratios
» The low-oxygen environment of the cell: Mitochondria between hypoxic and oxidative stress


  • Gnaiger E, Steinlechner-Maran R, Méndez G, Eberl T, Margreiter R (1995) Control of mitochondrial and cellular respiration by oxygen. J Bioenerg Biomembr 27:583-96. - »Bioblast link«
» First publication to establish high-resolution respirometry, based on the Oroboros Oxygraph.
» Standardized oxygen calibration (air and internal zero oxygen calibration).
» Automatic instrumental background correction of oxygen flux.
» Deconvolution of the oxygen signal for time resolution required in oxygen kinetics.
» Quality control of the limit of detection of oxygen flux at ±1 pmol O2·s-1·ml-1.


O2k-Manual

General

  • Worksheet O2k-Basic: This worksheet contains questions on the O2k and HRR addressed to participants of O2k-Basic Workshops. Answers to these questions are elaborated during the O2k-Workshop.