Gnaiger 2001 Respir Physiol

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Gnaiger E (2001) Bioenergetics at low oxygen: dependence of respiration and phosphorylation on oxygen and adenosine diphosphate supply. Respir Physiol 128:277-97.

» PMID: 11718759, Bioblast pdf

Gnaiger Erich (2001) Respir Physiol

Abstract: Oxygen limitation is generally considered as impairment of mitochondrial respiration under hypoxia and ischemia. Low intracellular oxygen levels under normoxia, however, imply mild oxygen limitation, provide protection from oxidative stress, and result from economical strategies for oxygen transport through the respiratory cascade to cytochrome c oxidase. Both perspectives relate to the critical oxygen pressure which inhibits mitochondrial respiration. Based on methodological considerations of oxygen kinetics and a presentation of high-resolution respirometry, mitochondrial oxygen affinities (1/p50) are reviewed with particular emphasis on the turnover effect under control of ADP, which increases the p2 in active states. ~P/O2 flux ratios are high even under severe oxygen limitation, as demonstrated by calorespirometry. Oxygen limitation reduces the uncoupled respiration observed under control by ADP, as shown by relationships derived between ~P/O2 flux ratios, respiratory control ratios, and ADP kinetics. Bioenergetics at low oxygen versus oxidative stress must be considered in the context of limitation of maximum aerobic activity, ischemia-reperfusion injury, mitochondrial signalling to apoptosis, and mitochondrial theories of ageing.

Keywords: Energy: Oxidative phosphorylation, Adenosine diphosphate kinetics, Adenosine diphosphate/O2 ratio; Hypoxia: Mitochondrial O2 kinetics, Mammals: Rat, Membrane permeability, Mitochondria: Heart, Liver

O2k-Network Lab: AT Innsbruck Gnaiger E

Gnaiger 2001 Respir Physiol New Fig10.jpg

Fig. 10 (modified): Opposite effects of ADP limitation and oxygen limitation on mitochondrial membrane potential, and LEAK oxygen flux, JO2,L. The LEAK state is obtained when total oxygen flux equals LEAK respiration. In all other respiratory states, total oxygen flux is the sum of LEAK oxygen flux and mechanistically coupled oxygen flux. A: ADP limitation of respiration at high oxygen levels in the transition from the OXPHOS state (P, saturating ADP) or active State 3 (high ADP) to the resting LEAK state, L (compare State 4), leads to an increase of membrane potential and exponential acceleration of the proton leak (heavy line). Because LEAK oxygen flux increases while total oxygen flux is reduced, the ATP yield (ADP/O2 flux ratio) declines to zero. Turnover-dependent proton leak increases the LEAK oxygen flux in the OXPHOS state but declines towards the LEAK state (Garlid et al 1993). Mitochondrial production of reactive oxygen species (ROS) increases with membrane potential towards the LEAK state, and ROS-linked electron bypass (electron leak) contributes minimally to LEAK oxygen flux at high oxygen (Gnaiger et al 2000). On the right, the decline of ~P/O2 flux ratios is shown in the transition from OXPHOS to LEAK. B: Oxygen limitation of respiration causes a reduction of membrane potential in the transition from ADP limitation at high oxygen, to intracellular conditions of low oxygen and low ADP, to finally severe oxygen limitation under hypoxia and anoxia. Potentially synergistic with the well documented membrane potential effect on LEAK flux, are the hypothetical effects of decreasing membrane permeability and suppression of ROS production under severe hypoxia, whereas intermediary levels of hypoxia may increase ROS production (modified from Gnaiger 2001; see original publication for further references).

* Discussion: Oxygen dependence of ROS production - are permeabilized fibers a valid model?.

Selected quotes

  • In the intracellular microenvironment, mitochondria are well separated from air-level oxygen pressure, and high rates of oxidative phosphorylation must be maintained near or at limiting oxygen levels in some tissues. On the other hand, mitochondria are routinely isolated and studied at unphysiologically high oxygen concentrations with limited additions of antioxidants, despite the fact that mitochondria in tissues are protected from oxidative stress by both low oxygen levels and complex defence systems against reactive oxygen species.
  • Mitochondrial p50 values are 5 to 20 times less than the half-saturation point of myoglobin, which then suggests that mitochondrial respiration operates at the edge of oxygen limitation under normoxic intracellular oxygen pressures (Gnaiger et al., 1998b).




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TermAbbreviationDescription
AerobicoxThe aerobic state of metabolism is defined by the presence of oxygen (air) and therefore the potential for oxidative reactions (ox) to proceed, particularly in oxidative phosphorylation (OXPHOS). Aerobic metabolism (with involvement of oxygen) is contrasted with anaerobic metabolism (without involvement of oxygen): Whereas anaerobic metabolism may proceed in the absence or presence of oxygen (anoxic or oxic conditions), aerobic metabolism is restricted to oxic conditions. Below the critical oxygen pressure, aerobic ATP production decreases.
AnaerobicAnaerobic metabolism takes place without the use of molecular oxygen, in contrast to aerobic metabolism. The capacity for energy assimilation and growth under anoxic conditions is the ultimate criterion for facultative anaerobiosis. Anaerobic metabolism may proceed not only under anoxic conditions or states, but also under hyperoxic and normoxic conditions (aerobic glycolysis), and under hypoxic and microxic conditions below the limiting oxygen pressure.
AnoxiaanoxIdeally the terms anoxia and anoxic (anox, without oxygen) should be restricted to conditions where molecular oxygen is strictly absent. Practically, effective anoxia is obtained when a further decrease of experimental oxygen levels does not elicit any physiological or biochemical response. The practical definition, therefore, depends on (i) the techiques applied for oxygen removal and minimizing oxygen diffusion into the experimental system, (ii) the sensitivity and limit of detection of analytical methods of measuring oxygen (O2 concentration in the nM range), and (iii) the types of diagnostic tests applied to evaluate effects of trace amounts of oxygen on physiological and biochemical processes. The difficulties involved in defining an absolute limit between anoxic and microxic conditions are best illustrated by a logarithmic scale of oxygen pressure or oxygen concentration. In the anoxic state (State 5), any aerobic type of metabolism cannot take place, whereas anaerobic metabolism may proceed under oxic or anoxic conditions.
Critical oxygen pressurepcThe critical oxygen pressure, pc, is defined as the partial oxygen pressure, pO2, below which aerobic catabolism (respiration or oxygen consumption) declines significantly. If anaerobic catabolism is activated simultaneously to compensate for lower aerobic ATP generation, then the limiting oxygen pressure, pl, is equal to the pc. In many cases, however, the pl is substantially lower than the pc.
HyperoxiahyperoxHyperoxia is defined as environmental oxygen pressure above the normoxic reference level. Cellular and intracellular hyperoxia is imposed on isolated cells and isolated mitochondria at air-level oxygen pressures which are higher compared to cellular and intracellular oxygen pressures under tissue conditions in vivo. Hyperoxic conditions may impose oxidative stress and may increase maximum aerobic performance.
HypoxiahypoxHypoxia (hypox) is defined as the state when insufficient O2 is available for respiration. This definition of hypoxia based on respiratory physiology is compared to environmental hypoxia defined as environmental oxygen pressures below the normoxic reference level.
Intracellular oxygenpO2,iPhysiological, intracellular oxygen pressure is significantly lower than air saturation under normoxia, hence respiratory measurements carried out at air saturation are effectively hyperoxic for cultured cells and isolated mitochondria.
Limiting oxygen pressureplThe limiting oxygen pressure, pl, is defined as the partial oxygen pressure, pO2, below which anaerobic catabolism is activated to contribute to total ATP generation. The limiting oxygen pressure, pl, may be substantially lower than the critical oxygen pressure, pc, below which aerobic catabolism (respiration or oxygen consumption) declines significantly.
MicroxiamicroxMicroxia (deep hypoxia) is obtained when trace amounts of O2 exert a stimulatory effect on respiration above the level where metabolism is switched to a purely anaerobic mode.
NormoxianormoxNormoxia is a reference state, frequently considered as air-level oxygen pressure at sea level (c. 20 kPa in water vapor saturated air) as environmental normoxia. Intracellular tissue normoxia is variable between organisms and tissues, and intracellular oxygen pressure is frequently well below air-level pO2 as a result of cellular (mainly mitochondrial) oxygen consumption and oxygen gradients along the respiratory cascade. Oxygen pressure drops from ambient normoxia of 20 kPa to alveolar normoxia of 13 kPa, while extracellular normoxia may be as low as 1 to 5 kPa in solid organs such as heart, brain, kidney and liver. Pericellular pO2 of cells growing in monolayer cell cultures may be hypoxic compared to tissue normoxia when grown in ambient normoxia (95 % air and 5 % CO2) and a high layer of culture medium causing oxygen diffusion limitation at high respiratory activity, but pericellular pO2 may be effectively hyperoxic in cells with low respiratory rate with a thin layer of culture medium (<2 mm). Intracellular oxygen levels in well-stirred suspended small cells (5 - 7 mm diameter; endothelial cells, fibroblasts) are close to ambient pO2 of the incubation medium, such that matching the experimental intracellular pO2 to the level of intracellular tissue normoxia requires lowering the ambient pO2 of the medium to avoid hyperoxia.
General
» Oxygen, dioxygen, O2
» Intracellular oxygen
» Oxygen pressure
» Oxygen solubility
» Gas pressure
» pascal
» Pressure
» Barometric pressure
» Concentration
Related keyword lists
» Keywords: Oxygen signal
» Keywords: Concentration and pressure

Publications: Tissue normoxia

 YearReferenceOrganismTissue;cellPreparationsStressDiseases
Stepanova 2020 Methods Cell Biol2020Stepanova A, Galkin A (2020) Measurement of mitochondrial H2O2 production under varying O2 tensions. Methods Cell Biol 155:273-93.MouseNervous systemIsolated mitochondriaOxidative stress;RONS
Ast 2019 Nat Metab2019Ast T, Mootha VK (2019) Oxygen and mammalian cell culture: are we repeating the experiment of Dr. Ox?. Nat Metab 1:858-860.
Keeley 2019 Physiol Rev2019Keeley TP, Mann GE (2019) Defining Physiological Normoxia for Improved Translation of Cell Physiology to Animal Models and Humans. Physiol Rev 99:161-234.
Stepanova 2018 J Neurochem2018Stepanova A, Konrad C, Manfredi G, Springett R, Ten V, Galkin A (2018) The dependence of brain mitochondria reactive oxygen species production on oxygen level is linear, except when inhibited by antimycin A. J Neurochem 148:731-45.MouseNervous systemIsolated mitochondriaIschemia-reperfusion
Oxidative stress;RONS
Stepanova 2018 J Cereb Blood Flow Metab2018Stepanova A, Konrad C, Guerrero-Castillo S, Manfredi G, Vannucci S, Arnold S, Galkin A (2018) Deactivation of mitochondrial complex I after hypoxia-ischemia in the immature brain. J Cereb Blood Flow Metab 39:1790-802.RatNervous systemIsolated mitochondriaHypoxia
Ischemia-reperfusion
Stuart 2018 Oxid Med Cell Longev2018Stuart JA, Fonseca JF, Moradi F, Cunningham C, Seliman B, Worsfold CR, Dolan S, Abando J, Maddalena LA (2018) How Supraphysiological Oxygen Levels in Standard Cell Culture Affect Oxygen-Consuming Reactions. Oxid Med Cell Longev 2018:8238459.
Stepanova 2017 J Cereb Blood Flow Metab2017Stepanova A, Kahl A, Konrad C, Ten V, Starkov AS, Galkin A (2017) Reverse electron transfer results in a loss of flavin from mitochondrial complex I: Potential mechanism for brain ischemia-reperfusion injury. J Cereb Blood Flow Metab 37:3649-58.MouseNervous systemIsolated mitochondriaIschemia-reperfusion
Harrison 2015 J Appl Physiol2015Harrison DK, Fasching M, Fontana-Ayoub M, Gnaiger E (2015) Cytochrome redox states and respiratory control in mouse and beef heart mitochondria at steady-state levels of hypoxia. J Appl Physiol 119:1210-8.Mouse
Bovines
HeartIsolated mitochondriaHypoxia
Carreau 2011 J Cell Mol Med2011Carreau A, El Hafny-Rahbi B, Matejuk A, Grillon C, Kieda C (2011) Why is the partial oxygen pressure of human tissues a crucial parameter? Small molecules and hypoxia. J Cell Mol Med 15:1239-53.
Aragones 2009 Cell Metab2009Aragones J, Fraisl P, Baes M, Carmeliet P (2009) Oxygen sensors at the crossroad of metabolism. Cell Metab 9:11-22.
Pettersen 2005 Cell Prolif2005Pettersen EO, Larsen LH, Ramsing NB, Ebbesen P (2005) Pericellular oxygen depletion during ordinary tissue culturing, measured with oxygen microsensors. Cell Prolif 38:257-67.
Gnaiger 2003 Adv Exp Med Biol2003Gnaiger E (2003) Oxygen conformance of cellular respiration. A perspective of mitochondrial physiology. Adv Exp Med Biol 543:39-55.Human
Rat
Heart
Liver
Endothelial;epithelial;mesothelial cell
Fibroblast
Intact cells
Permeabilized cells
Permeabilized tissue
Isolated mitochondria
Oxidase;biochemical oxidation
Gnaiger 2001 Respir Physiol2001Gnaiger E (2001) Bioenergetics at low oxygen: dependence of respiration and phosphorylation on oxygen and adenosine diphosphate supply. Respir Physiol 128:277-97.Human
Rat
Heart
Liver
Endothelial;epithelial;mesothelial cell
HUVEC
Intact cells
Isolated mitochondria
Oxidative stress;RONS
Gnaiger 2000 Proc Natl Acad Sci U S A2000Gnaiger E, Méndez G, Hand SC (2000) High phosphorylation efficiency and depression of uncoupled respiration in mitochondria under hypoxia. Proc Natl Acad Sci U S A 97:11080-5.Rat
Artemia
Crustaceans
LiverIsolated mitochondria
Gnaiger 1998 J Exp Biol1998Gnaiger E, Lassnig B, Kuznetsov AV, Rieger G, Margreiter R (1998) Mitochondrial oxygen affinity, respiratory flux control, and excess capacity of cytochrome c oxidase. J Exp Biol 201:1129-39.Human
Rat
Heart
Liver
Endothelial;epithelial;mesothelial cell
HUVEC
Isolated mitochondria
Enzyme
Oxidase;biochemical oxidation
Intact cells
Gnaiger 1998 Biochim Biophys Acta1998Gnaiger E, Lassnig B, Kuznetsov AV, Margreiter R (1998) Mitochondrial respiration in the low oxygen environment of the cell: Effect of ADP on oxygen kinetics. Biochim Biophys Acta 1365:249-54.RatHeart
Liver
Isolated mitochondria
Gnaiger 1995 J Bioenerg Biomembr1995Gnaiger 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.Human
Rat
Liver
Endothelial;epithelial;mesothelial cell
HUVEC
Isolated mitochondria
Intact cells
Gnaiger 1993 Transitions1993Gnaiger E (1993) Homeostatic and microxic regulation of respiration in transitions to anaerobic metabolism. In: The vertebrate gas transport cascade: Adaptations to environment and mode of life. Bicudo JEPW (ed), CRC Press, Boca Raton, Ann Arbor, London, Tokyo:358-70.Reptiles
Fishes
Crustaceans
Annelids
Intact organism
Gnaiger 1991 Soc Exp Biol Seminar Series1991Gnaiger E (1991) Animal energetics at very low oxygen: Information from calorimetry and respirometry. In: Strategies for gas exchange and metabolism. Woakes R, Grieshaber M, Bridges CR (eds), Soc Exp Biol Seminar Series 44, Cambridge Univ Press, London:149-71.AnnelidsIntact organism
Gnaiger 1983 J Exp Zool1983Gnaiger E (1983) Heat dissipation and energetic efficiency in animal anoxibiosis. Economy contra power. J Exp Zool 228:471-90.Annelids
Molluscs
Skeletal muscleIntact organism
Abstracts: Tissue normoxia
 YearReferenceOrganismTissue;cellPreparationsStressDiseases
Gnaiger 2018 AussieMit2018Komlodi Timea, Sobotka Ondrej, Doerrier Carolina, Gnaiger Erich (2018) Mitochondrial H2O2 production is low under tissue normoxia but high at in-vitro air-level oxygen pressure - comparison of LEAK and OXPHOS states. AussieMit 2018 Melbourne AU.Mouse
Saccharomyces cerevisiae
Heart
Nervous system
Isolated mitochondria
Intact cells
Oxidative stress;RONS
Hypoxia
Sobotka 2018 MiP20182018
Ondrej Sobotka
Measurement of ROS production under hypoxia and unexpected methodological pitfalls of Amplex UltraRed assay.
Mouse
Saccharomyces cerevisiae
Heart
Nervous system
Isolated mitochondriaHypoxia
Komlodi 2017 MiP20172017
Timea Komlodi
H2O2 production under hypoxia in brain and heart mitochondria: does O2 concentration matter?
MouseHeart
Nervous system
Isolated mitochondriaOxidative stress;RONS
Hypoxia


Cited by

Gnaiger E (2021) Bioenergetic cluster analysis – mitochondrial respiratory control in human fibroblasts. MitoFit Preprints 2021.8.


Gnaiger E (2021) Bioenergetic cluster analysis – mitochondrial respiratory control in human fibroblasts. MitoFit Preprints 2021.8. - »Bioblast link«
  • Cardoso et al (2021) Magnesium Green for fluorometric measurement of ATP production does not interfere with mitochondrial respiration. MitoFit Preprints 2021.1. doi:10.26124/mitofit:2021-0001
  • Komlodi et al (2021) Simultaneous measurement of respiration and redox state of the Coenzyme Q pool in mitochondrial preparations. MitoFit Preprints 2021.2.v3 doi:10.26124/mitofit:2021-0002.v3
  • Komlódi T, Sobotka O, Gnaiger E (2021) Facts and artefacts on the oxygen dependence of hydrogen peroxide production using Amplex UltraRed. MitoFit Preprints 2021 (in prep). - Bioblast link
  • Komlodi T et al (2021) Oxygen dependence of hydrogen peroxide production using Amplex UltraRed in yeast cells, isolated mitochondria, and permeabilized cells. MitoFit Preprints 2021 (in prep).
  • Komlodi et al (2021) Hydrogen peroxide production, mitochondrial membrane potential and the redox state of the coenzyme Q measured at tissue normoxia and experimental hyperoxia in heart mitochondria. MitoFit Preprints 2021 (in prep)
Gnaiger 2020 BEC MitoPathways


Gnaiger E (2020) Mitochondrial pathways and respiratory control. An introduction to OXPHOS analysis. 5th ed. Bioenerg Commun 2020.2:112 pp. doi:10.26124/bec:2020-0002
Gnaiger Erich et al ― MitoEAGLE Task Group (2020) Mitochondrial physiology. Bioenerg Commun 2020.1.


Gnaiger E et al ― MitoEAGLE Task Group (2020) Mitochondrial physiology. Bioenerg Commun 2020.1. doi:10.26124/bec:2020-0001.v1.


Labels: MiParea: Respiration, Instruments;methods, Comparative MiP;environmental MiP 

Stress:Oxidative stress;RONS  Organism: Human, Rat  Tissue;cell: Heart, Liver, Endothelial;epithelial;mesothelial cell, HUVEC  Preparation: Isolated mitochondria, Intact cells 

Regulation: ADP, Coupling efficiency;uncoupling, Oxygen kinetics, Threshold;excess capacity  Coupling state: OXPHOS 

HRR: Oxygraph-2k, TIP2k 

ATP, Steady state, Tissue normoxia, BEC 2020.1, BEC 2020.2, MitoFit 2021 MgG, MitoFit 2021 CoQ, MitoFit 2021 AmR-O2, MitoFit 2021 AmR, MitoFit 2021 Tissue normoxia, MitoFit 2021 BCA