Cookies help us deliver our services. By using our services, you agree to our use of cookies. More information

IOC recommended reading

From Bioblast

                



IOC recommended reading

COST Action MitoEAGLE

MitoEAGLE 'Mitochondrial physiology' BEC 2020.1

  • Gnaiger Erich et al ― MitoEAGLE Task Group (2020) Mitochondrial physiology. Bioenerg Commun 2020.1. doi:10.26124/bec:2020-0001.v1. - »Bioblast link«


O2k-Workshops.png
O2k-Publications in the MiPMap


Workshop material

  • Gnaiger Erich et al ― MitoEAGLE Task Group (2020) Mitochondrial physiology. Bioenerg Commun 2020.1. doi:10.26124/bec:2020-0001.v1. - »Bioblast link«
  • Gnaiger E (2020) Mitochondrial pathways and respiratory control. An introduction to OXPHOS analysis. 5th ed. Bioenerg Commun 2020.2. - »Bioblast link« - A new edition with updated terminology and more ..
  • Baglivo E, Cardoso LHD, Cecatto C, Gnaiger E (2022) Statistical analysis of instrumental reproducibility as internal quality control in high-resolution respirometry. Bioenerg Commun 2022.8. »Bioblast link«
  • 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«
  • Komlódi T, Sobotka O, Gnaiger E (2021) Facts and artefacts on the oxygen dependence of hydrogen peroxide production using Amplex UltraRed. Bioenerg Commun 2021.4 »Bioblast pdf«


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«
» Introduction to harmonized reference protocols 1 and 2 (SUIT-001 and 002) for mitochondrial preparation.
» Introduction to SUIT protocols with living cells (SUIT-003).
» Optimum digitonin concentration test (SUIT-010).
» Complex IV activity and chemical background correction.
» Introduction to O2 kinetics.


  • 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 (2020) Mitochondrial pathways and respiratory control. An introduction to OXPHOS analysis. 5th ed. BEC 2020.2. - »Bioblast link«
» A hard copy of this monograph on mitochondrial physiology will be included in the workshop material. The Blue Book is available online 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.
» Living cells, permeabilized cells and tissue, or isolated mitochondria?




OXPHOS ROUTINE ETS LEAK ROX


O2k-FluoRespirometer

O2k-FluoRespirometry

  • Komlódi T, Sobotka O, Gnaiger E (2021) Facts and artefacts on the oxygen dependence of hydrogen peroxide production using Amplex UltraRed. Bioenerg Commun 2021.4 »Bioblast pdf«
  • Cardoso LHD, Doerrier C, Gnaiger E (2021) Magnesium Green for fluorometric measurement of ATP production does not interfere with mitochondrial respiration. Bioenerg Commun 2021.1 »Bioblast pdf«
  • Komlodi T, Sobotka O, Krumschnabel G, Bezuidenhout N, Hiller E, Doerrier C, Gnaiger E (2018) Comparison of mitochondrial incubation media for measurement of respiration and hydrogen peroxide production. Methods Mol Biol 1782:137-55. »Bioblast pdf«
  • 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

  • Zdrazilova L, Hansikova H, Gnaiger E (2022) Comparable respiratory activity in attached and suspended human fibroblasts. PLoS ONE 17:e0264496 - »Bioblast link«
» Respirometry with living cells.
» Repeatability and reproducibility.


  • Gnaiger E (2009) Capacity of oxidative phosphorylation in human skeletal muscle. New perspectives of mitochondrial physiology. Int J Biochem Cell Biol 41:1837-45. - »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 biopsies 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.