Gnaiger 2000 Proc Natl Acad Sci U S A

Gnaiger 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. https://doi.org/10.1073/pnas.97.20.11080

» PMID: 11005877 Open Access

Gnaiger Erich, Mendez Gabriela, Hand Steven C (2000) Proc Natl Acad Sci U S A

Abstract: Mitochondria are confronted with low oxygen levels in the microenvironment within tissues; yet, isolated mitochondria are routinely studied under air-saturated conditions that are effectively hyperoxic, increase oxidative stress, and may impair mitochondrial function. Under hypoxia, on the other hand, respiration and ATP supply are restricted. Under these conditions of oxygen limitation, any compromise in the coupling of oxidative phosphorylation to oxygen consumption could accentuate ATP depletion, leading to metabolic failure. To address this issue, we have developed the approach of oxygen-injection microcalorimetry and ADP-injection respirometry for evaluating mitochondrial function at limiting oxygen supply. Whereas phosphorylation efficiency drops during ADP limitation at high oxygen levels, we show here that oxidative phosphorylation is more efficient at low oxygen than at air saturation, as indicated by higher ratios of ADP flux to total oxygen flux at identical submaximal rates of ATP synthesis. At low oxygen, the proton leak and uncoupled respiration are depressed, thus reducing maintenance energy expenditure. This indicates the importance of low intracellular oxygen levels in avoiding oxidative stress and protecting bioenergetic efficiency.

• O2k-Network Lab: AT Innsbruck Oroboros, AT Innsbruck Gnaiger E, US LA Baton Rouge Hand SC

Selected quotes

Atmospheric oxygen levels were probably 0.1 % of the present when mitochondria became associated with cells early during evolution (1-4). Such low-oxygen conditions persist in extreme environments (5-8), and oxygen pessures as low as 0.3-0.4 kPa (2-3 mmHg) are observed in the intracellular microenvironment of mitochondria in tissues under normoxia (9-13). Even so, in typical studies with isolated mitochondria, these organelles are artificially exposed to the high partial pressure of oxygen at air saturation (c. 20 kPa), despite the fact that this condition is effectively hyperoxic, rarely physiological and increases oxidative stress (14, 15).

From the present study, it can be inferred that air-saturation as a reference condition is questionable for evaluating mitochondrial efficiency. If representative measures of performance are to be obtained for mitochondria and even cultured cells (42, 44), then oxygen tension should be shifted away from these non-physiological levels to minimize the risk of oxidative stress.

Furthermore, the high efficiency of oxidative phosphorylation at low oxygen emphasizes that even trace amounts of oxygen can make a vital energetic contribution when ATP limitation threatens cellular survival under severe hypoxia encountered at high altitude, in aquatic habitats, and during pathological states of ischemia.

Further details

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«

» OXPHOS-coupling efficiency

» ETS coupling efficiency

Keywords: Oxia terms

Bioblast links: Hypoxia, normoxia, hyperoxia - >>>>>>> - Click on [Expand] or [Collapse] - >>>>>>>

Term	Abbreviation	Description
Aerobic	ox	The 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.
Anaerobic		Anaerobic 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.
Anoxia	anox	Ideally 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 pressure	p_c	The critical oxygen pressure, p_c, is defined as the partial oxygen pressure, p_O2, 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, p_l, is equal to the p_c. In many cases, however, the p_l is substantially lower than the p_c.
Hyperoxia	hyperox	Hyperoxia 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.
Hypoxia	hypox	Hypoxia (hypox) is defined in respiratory physiology as the state when insufficient O₂ is available for respiration, compared to environmental hypoxia defined as environmental oxygen pressures below the normoxic reference level. Three major categories of hypoxia are (1) environmental hypoxia, (2) physiological tissue hypoxia in hyperactivated states (e.g. at V_O₂max) with intracellular oxygen demand/supply balance at steady state in tissues at environmental normoxia, compared to tissue normoxia in physiologically balanced states, and (3) pathological tissue hypoxia including ischemia and stroke, anaemia, chronic heart disease, chronic obstructive pulmonary disease, severe COVID-19, and obstructive sleep apnea. Pathological hypoxia leads to tissue hypoxia and heterogenous intracellular anoxia. Clinical oxygen treatment ('environmental hyperoxia') may not or only partially overcome pathological tissue hypoxia.
Intracellular oxygen	p_O2,i	Physiological, 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 pressure	p_l	The limiting oxygen pressure, p_l, is defined as the partial oxygen pressure, p_O2, below which anaerobic catabolism is activated to contribute to total ATP generation. The limiting oxygen pressure, p_l, may be substantially lower than the critical oxygen pressure, p_c, below which aerobic catabolism (respiration or oxygen consumption) declines significantly.
Microxia	microx	Microxia (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.
Normoxia	normox	Normoxia 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 p_O₂ 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 p_O₂ 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 p_O₂ 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 p_O₂ of the incubation medium, such that matching the experimental intracellular p_O₂ to the level of intracellular tissue normoxia requires lowering the ambient p_O₂ 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

	Year	Reference	Organism	Tissue;cell	Preparations	Stress
Donnelly 2022 BEC	2022	Donnelly C, Schmitt S, Cecatto C, Cardoso LHD, Komlódi T, Place N, Kayser B, Gnaiger E (2022) The ABC of hypoxia – what is the norm. Bioenerg Commun 2022.12.v2. https://doi.org/10.26124/bec:2022-0012.v2				Oxidative stress;RONS Hypoxia
Donnelly 2022 MitoFit Hypoxia	2022	Donnelly C, Schmitt S, Cecatto C, Cardoso LHD, Komlodi T, Place N, Kayser B, Gnaiger E (2022) The ABC of hypoxia – what is the norm. https://doi.org/10.26124/mitofit:2022-0025.v2 — 2022-11-14 published in Bioenerg Commun 2022.12.				Oxidative stress;RONS Hypoxia
DiProspero 2021 Toxicol In Vitro	2021	DiProspero TJ, Dalrymple E, Lockett MR (2021) Physiologically relevant oxygen tensions differentially regulate hepatotoxic responses in HepG2 cells. https://doi.org/10.1016/j.tiv.2021.105156		Liver	Intact cells	Hypoxia
Stepanova 2020 Methods Cell Biol	2020	Stepanova A, Galkin A (2020) Measurement of mitochondrial H₂O₂ production under varying O₂ tensions. https://doi.org/10.1016/bs.mcb.2019.12.008	Mouse	Nervous system	Isolated mitochondria	Oxidative stress;RONS
Keeley 2019 Physiol Rev	2019	Keeley TP, Mann GE (2019) Defining physiological normoxia for improved translation of cell physiology to animal models and humans. https://doi.org/10.1152/physrev.00041.2017
Ast 2019 Nat Metab	2019	Ast T, Mootha VK (2019) Oxygen and mammalian cell culture: are we repeating the experiment of Dr. Ox? Nat Metab 1:858-860.
Stepanova 2018 J Neurochem	2018	Stepanova 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.	Mouse	Nervous system	Isolated mitochondria	Ischemia-reperfusion Oxidative stress;RONS
Stuart 2018 Oxid Med Cell Longev	2018	Stuart 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 2018 J Cereb Blood Flow Metab	2018	Stepanova 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.	Rat	Nervous system	Isolated mitochondria	Hypoxia Ischemia-reperfusion
Stepanova 2017 J Cereb Blood Flow Metab	2017	Stepanova 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.	Mouse	Nervous system	Isolated mitochondria	Ischemia-reperfusion
Harrison 2015 J Appl Physiol	2015	Harrison 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. https://doi.org/10.1152/japplphysiol.00146.2015	Mouse Bovines	Heart	Isolated mitochondria	Hypoxia
Carreau 2011 J Cell Mol Med	2011	Carreau 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. https://doi.org/10.1111/j.1582-4934.2011.01258.x
Richardson 2006 J Physiol	2006	Richardson RS, Duteil S, Wary C, Wray DW, Hoff J, Carlier PG (2006) Human skeletal muscle intracellular oxygenation: the impact of ambient oxygen availability. https://doi.org/10.1113/jphysiol.2005.102327	Human	Skeletal muscle		Hypoxia
Pettersen 2005 Cell Prolif	2005	Pettersen 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 Biol	2003	Gnaiger E (2003) Oxygen conformance of cellular respiration. A perspective of mitochondrial physiology. https://doi.org/10.1007/978-1-4419-8997-0_4	Human Rat	Heart Liver Endothelial;epithelial;mesothelial cell Fibroblast	Intact cells Permeabilized cells Permeabilized tissue Isolated mitochondria Oxidase;biochemical oxidation
Gnaiger 2001 Respir Physiol	2001	Gnaiger E (2001) Bioenergetics at low oxygen: dependence of respiration and phosphorylation on oxygen and adenosine diphosphate supply. https://doi.org/10.1016/S0034-5687(01)00307-3	Human Rat	Heart Liver Endothelial;epithelial;mesothelial cell HUVEC	Intact cells Isolated mitochondria	Oxidative stress;RONS
Gnaiger 2000 Proc Natl Acad Sci U S A	2000	Gnaiger 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. https://doi.org/10.1073/pnas.97.20.11080	Rat Artemia Crustaceans	Liver	Isolated mitochondria
Gnaiger 1998 J Exp Biol	1998	Gnaiger E, Lassnig B, Kuznetsov AV, Rieger G, Margreiter R (1998) Mitochondrial oxygen affinity, respiratory flux control, and excess capacity of cytochrome c oxidase. https://doi.org/10.1242/jeb.201.8.1129	Human Rat	Heart Liver Endothelial;epithelial;mesothelial cell HUVEC	Isolated mitochondria Enzyme Oxidase;biochemical oxidation Intact cells
Gnaiger 1998 Biochim Biophys Acta	1998	Gnaiger 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. https://doi.org/10.1016/S0005-2728(98)00076-0	Rat	Heart Liver	Isolated mitochondria
Gnaiger 1995 J Bioenerg Biomembr	1995	Gnaiger E, Steinlechner-Maran R, Méndez G, Eberl T, Margreiter R (1995) Control of mitochondrial and cellular respiration by oxygen. https://doi.org/10.1007/BF02111656	Human Rat	Liver Endothelial;epithelial;mesothelial cell HUVEC	Isolated mitochondria Intact cells
Gnaiger 1993 Transitions	1993	Gnaiger 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 Series	1991	Gnaiger 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.	Annelids		Intact organism
Gnaiger 1983 J Exp Zool	1983	Gnaiger E (1983) Heat dissipation and energetic efficiency in animal anoxibiosis. Economy contra power. J Exp Zool 228:471-90.	Annelids Molluscs	Skeletal muscle	Intact organism

Abstracts: Tissue normoxia

	Year	Reference	Organism	Tissue;cell	Preparations	Stress
Donnelly 2022 Abstract Bioblast	2022	2.1. «10+5» Donnelly Chris, Schmitt S, Cecatto C, Cardoso L, Komlodi T, Place N, Kayser B, Gnaiger E (2022) The ABC of hypoxia – what is the norm. Bioblast 2022: BEC Inaugural Conference. In: https://doi.org/10.26124/bec:2022-0001 »MitoFit Preprint«				Oxidative stress;RONS Hypoxia
Gnaiger 2018 AussieMit	2018	Komlodi 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 MiP2018	2018	Measurement of ROS production under hypoxia and unexpected methodological pitfalls of Amplex UltraRed assay.	Mouse Saccharomyces cerevisiae	Heart Nervous system	Isolated mitochondria	Hypoxia
Komlodi 2017 MiP2017	2017	H₂O₂ production under hypoxia in brain and heart mitochondria: does O₂ concentration matter?	Mouse	Heart Nervous system	Isolated mitochondria	Oxidative stress;RONS Hypoxia

Citations

Cited by

Gnaiger E (2024) Addressing the ambiguity crisis in bioenergetics and thermodynamics. MitoFit Preprints 2024.3. https://doi.org/10.26124/mitofit:2024-0003

Cardoso et al (2021) Magnesium Green for fluorometric measurement of ATP production does not interfere with mitochondrial respiration. Bioenerg Commun 2021.1. doi:10.26124/bec:2021-0001

Gnaiger E (2020) Mitochondrial pathways and respiratory control. An introduction to OXPHOS analysis. 5^th ed. Bioenerg Commun 2020.2. https://doi.org/10.26124/bec:2020-0002

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

70 articles in PubMed (2024-05-01) https://pubmed.ncbi.nlm.nih.gov/11005877/

Labels: MiParea: Respiration, mt-Biogenesis;mt-density, mt-Structure;fission;fusion, Comparative MiP;environmental MiP, Developmental biology

Organism: Rat, Artemia, Crustaceans Tissue;cell: Liver Preparation: Isolated mitochondria

Regulation: ADP, ATP, Coupling efficiency;uncoupling, Oxygen kinetics Coupling state: LEAK, OXPHOS Pathway: S HRR: Oxygraph-2k, TIP2k

Ambiguity crisis, Tissue normoxia, ATP, Steady state, BEC 2020.1, BEC 2020.2, MitoFit 2021 MgG, MitoFit2022rTCA, MitoFit2022Hypoxia, Gnaiger 2024 MitoFit