IOC recommended reading: Difference between revisions
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**The low-oxygen environment of the cell: Mitochondria between hypoxic and oxidative stress | **The low-oxygen environment of the cell: Mitochondria between hypoxic and oxidative stress | ||
*'''Scandurra FM, Gnaiger E (2010)''' Cell respiration under hypoxia: Facts and artefacts in mitochondrial oxygen kinetics. Adv Exp Med Biol 662: 7-25. [[Scandurra 2010 Adv Exp Med Biol|''' | *'''Scandurra FM, Gnaiger E (2010)''' Cell respiration under hypoxia: Facts and artefacts in mitochondrial oxygen kinetics. Adv Exp Med Biol 662: 7-25. [[Scandurra 2010 Adv Exp Med Biol|'''ยปOpen Access''']] ย | ||
**Isolated mitochondria and intact cells ย | **Isolated mitochondria and intact cells ย | ||
**Nanomolar resolution of oxygen concentration ย | **Nanomolar resolution of oxygen concentration ย | ||
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**Respiration per million cells, per citrate synthase, per mg protein, or per cytochrome c oxidase activity. | **Respiration per million cells, per citrate synthase, per mg protein, or per cytochrome c oxidase activity. | ||
*[[O2k-Publications|O2k-Publications]] | *[[O2k-Publications|'''O2k-Publications''']] | ||
*[[O2k-Protocols|O2k-Protocols]] | *[[O2k-Protocols|'''O2k-Protocols''']] | ||
*[[Respiratory states|Respiratory states]] | *[[Respiratory states|'''Respiratory states''']] | ||
== O2k-Workshop TPP/ISE == | == O2k-Workshop TPP/ISE == | ||
*O2k-MultiSensor System with Ion Selective Electrodes (ISE) | *O2k-MultiSensor System with Ion Selective Electrodes (ISE) [[MiPNet15.03]] | ||
*Determination of membrane potential using TPP+ and an ion selective electrode (ISE) | *Determination of membrane potential using TPP+ and an ion selective electrode (ISE) [[MiPNet14.05]] | ||
*Calculation of mitochondrial membrane potential from measurements with a TPP electrode | *[[Calculation of mitochondrial membrane potential from measurements with a TPP electrode]] | ||
*Rottenberg H (1984) Membrane potential and surface potential in mitochondria: uptake and binding of lipophilic cations. J Membr Biol 81: 127-138. | *Rottenberg H (1984) Membrane potential and surface potential in mitochondria: uptake and binding of lipophilic cations. J Membr Biol 81: 127-138. |
Revision as of 11:03, 26 June 2014
General reading
- ยปWorksheet 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.
- Gnaiger E (2012) Mitochondrial pathways and respiratory control. An introduction to OXPHOS analysis. 3rd ed. Mitochondr Physiol Network 17.18. OROBOROS MiPNet Publications, Innsbruck: 64 pp. ยปOpen Access
- The blue book: OXPHOS analysis; CI linked, CII linked, CI+II linked pathways, respiratory states, abbreviations
- 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. ยปOpen Access
- Introduction to high-resolution respirometry (HRR), SUIT protocols and flux control ratios.
- 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: 327-352. ยปOpen Access
- Methodological introduction to HRR, with focus on Polarographic oxygen sensor and traditional oxygraphy
- Dramatically different: High-resolution respirometry - the Oxygraph-2k
- 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 (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?
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. ยปAbstract
- Respirometry with permeabilized fibres and isolated mitochondria
- Convergent CI+II electron input and OXPHOS capacity
- Tissue-OXPHOS capacity in human permeabilized muscle fibres and isolated mitochondria
- Tissue-OXPHOS capacity and functional diversity
- Gnaiger E (2001) Bioenergetics at low oxygen: dependence of respiration and phosphorylation on oxygen and adenosine diphosphate supply. Respir Physiol 128: 277-297. โ A detailed introduction into high-resolution respirometry with particular emphasis on kinetics and measurements at low oxygen. ยปAbstract
- 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
- Scandurra FM, Gnaiger E (2010) Cell respiration under hypoxia: Facts and artefacts in mitochondrial oxygen kinetics. Adv Exp Med Biol 662: 7-25. ยปOpen Access
- 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: 431-442. ยปAbstract
- Isolated mitochondria and permeabilized muscle fibers, MiR05:
- Optimization of mitochondrial cold storage โข Mitochondrial respiration medium, MiR05 โข 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: 115-123. ยปAbstract
- Intact cells
- Cytochrome c oxidase
- Cytochrome c test
- Respiration per million cells, per citrate synthase, per mg protein, or per cytochrome c oxidase activity.
O2k-Workshop 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: 127-138.