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Difference between revisions of "Cell ergometry"

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{{MitoPedia
{{MitoPedia
|abbr=n.a.
|description=Biochemical '''cell ergometry''' aims at measurement of ''J''<sub>O<sub>2</sub>max</sub> (compare ''V''<sub>O<sub>2</sub>max</sub> or ''V''<sub>O<sub>2</sub>peak</sub> in exercise ergometry of humans and animals) of cell respiration linked to phosphorylation of ADP to ATP. The corresponding [[OXPHOS capacity]] is based on saturating concentrations of ADP, [ADP], and inorganic phosphate [Pi] available to the mitochondria. This is metabolically opposite to uncoupling of respiration, which yields [[ET capacity]].  The OXPHOS state can be established experimentally by selective [[permeabilized cells |permeabilization of cell membranes]] with maintenance of intact mitochondria, titrations of ADP and P<sub>i</sub> to evaluate kinetically saturating conditions, and establishing fuel substrate combinations which reconstitute physiological [[TCA cycle]] function. Uncoupler titrations are applied to determine the apparent ET-pathway excess over OXPHOS capacity ([[E-P control efficiency |''E-P'' control efficiency) and to calculate the [[P-L control efficiency |''P-L'' control efficiency]] ''j<sub>P-L</sub>'' and [[E-L coupling efficiency |''E-L'' coupling efficiency]] ''j<sub>E-L</sub>''. These normalized flux ratios are the basis to calculate the ergometric or [[ergodynamic efficiency]], ''ε'' = ''j'' · ''f'', where ''f'' is the normalized force ratio.
|description=Biochemical '''cell ergometry''' aims at measurement of ''J''<sub>O2max</sub> (compare ''V''<sub>O2max</sub> or ''V''<sub>O2peak</sub> in exercise ergometry of humans and animals) of cell respiration linked to phosphorylation of ADP to ATP. The corresponding [[OXPHOS capacity]] is based on saturating concentrations of ADP, [ADP]*, and inorganic phosphate, [Pi]*, available to the mitochondria. This is metabolically opposite to uncoupling respiration, which yields [[ETS capacity]].  The OXPHOS state can be established experimentally by selective [[permeabilized cells |permeabilization of cell membranes]] with maintenance of intact mitochondria, titrations of ADP and P<sub>i</sub> to evaluate kinetically saturating conditions, and establishing fuel substrate combinations which reconstitute physiological [[TCA cycle]] function. Uncoupler titrations are applied to determine the apparent ETS excess over OXPHOS capacity and to calculate [[OXPHOS coupling efficiency |OXPHOS-]] and [[ETS coupling efficiency |ETS coupling efficiencies]], ''j<sub>≈P</sub>'' and ''j<sub>≈E</sub>''. These normalized flux ratios are the basis to calculate the ergometric or [[ergodynamic efficiency]], ''ε'' = ''j'' · ''f'', where ''f'' is the normalized force ratio.


» [[Cell_ergometry#Cell_ergometry_and_OXPHOS|'''MiPNet article''']]
» [[Cell_ergometry#Cell_ergometry_and_OXPHOS|'''MiPNet article''']]
|info=[[Gnaiger 2014 MitoPathways]]
|info=[[Gnaiger 2020 BEC MitoPathways]], [[Oxygen flux]]
}}
{{MitoPedia methods
|mitopedia method=Respirometry
}}
{{MitoPedia topics
|mitopedia topic=Respiratory state
}}
}}
__TOC__
__TOC__
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|title=Gnaiger E (2015) Cell ergometry and OXPHOS. Mitochondr Physiol Network 2015-01-18.
|title=Gnaiger E (2015) Cell ergometry and OXPHOS. Mitochondr Physiol Network 2015-01-18.
|info=
|info=
|authors=OROBOROS
|authors=Oroboros
|year=2015
|year=2015
|journal=MiPNet
|journal=MiPNet
|abstract=Spiroergometry on the organismic level is compared to cell ergometry as OXPHOS analysis on the cellular level.
|abstract=Spiroergometry on the organismic level is compared to cell ergometry as OXPHOS analysis on the cellular level.
|mipnetlab=AT Innsbruck Gnaiger E
|mipnetlab=AT Innsbruck Gnaiger E
}}
{{Labeling
|topics=Coupling efficiency;uncoupling
|couplingstates=LEAK, OXPHOS, ETS
|instruments=Theory
}}
}}
__TOC__
__TOC__
[[File:Cell ergometry.jpg|960px]]
[[File:Cell ergometry.pdf|960px]]
 
 
<big><big>'''Figure 1: [[Coupling control protocol]] in the intact cell'''</big></big>
 


== Spiroergometry ==
== Spiroergometry ==
''V''<sub>O2max</sub> or ''V''<sub>O2peak</sub> in cycle or treadmill '''spiroergometry''' is expressed in units of [ml O<sub>2</sub>·min<sup>-1</sup>·kg<sup>-1</sup>] body mass. 1 ml oxygen at STPD is equivalent to 22.392 mmol O<sub>2</sub>. Therefore, multiply by 1000/(22.392·60)=0.744 to convert ''V''<sub>O2peak</sub> to ''J''<sub>O2peak</sub> expressed in ''SI'' units [nmol·s<sup>-1</sup>·g<sup>-1</sup>]:
:::: ''V''<sub>O<sub>2</sub>max</sub> or ''V''<sub>O<sub>2</sub>peak</sub> in cycle or treadmill '''spiroergometry''' is expressed in units of [mL O<sub>2</sub>·min<sup>-1</sup>·kg<sup>-1</sup>] body mass. 1 mL oxygen at [[STPD]] is equivalent to 22.392 mmol O<sub>2</sub>. Therefore, multiply by 1000/(22.392·60)=0.744 to convert ''V''<sub>O<sub>2</sub>max</sub> to ''J''<sub>O<sub>2</sub>max</sub> expressed in SI units [nmol·s<sup>-1</sup>·g<sup>-1</sup>]:
  <big>1 ml O<sub>2</sub>·min<sup>-1</sup>·kg<sup>-1</sup> = 0.744 µmol·s<sup>-1</sup>·kg<sup>-1</sup></big>
  <big>1 mL O<sub>2</sub>·min<sup>-1</sup>·kg<sup>-1</sup> = 0.744 µmol·s<sup>-1</sup>·kg<sup>-1</sup></big>


''V''<sub>O2peak</sub> (''J''<sub>O2peak</sub>) typically declines from 70 to 25 ml O<sub>2</sub>·min<sup>-1</sup>·kg<sup>-1</sup> (50 to 20 µmol·s<sup>-1</sup>·kg<sup>-1</sup>) in the range of healthy trained to obese untrained humans.
:::: ''V''<sub>O<sub>2</sub>max</sub> (''J''<sub>O<sub>2</sub>max</sub>) typically declines from 70 to 25 mL O<sub>2</sub>·min<sup>-1</sup>·kg<sup>-1</sup> (50 to 20 µmol·s<sup>-1</sup>·kg<sup>-1</sup>) in the range of healthy trained to obese untrained humans.


== Keywords ==
::::* Expand Bioblast links to '''Cell ergometry'''
{{Template:Keywords: Coupling control}}


== Cell ergometry: intact cells ==
=== Respiratory coupling states in intact cells ===
:: [[Image:R.jpg|link=ROUTINE respiration|ROUTINE]] [[ROUTINE respiration]], ''R'' = ''R´''-ROX
:::::::: [[Image:R-L.jpg|50 px|link=Free ROUTINE activity |Free ROUTINE activity]] [[Free ROUTINE activity]], ''≈R'' = ''R-L''
:: [[Image:E.jpg|link=ETS capacity|ETS]] [[ETS capacity]], ''E'' = ''E´''-ROX
:::::::: [[Image:E-L.jpg|50 px|link=Free ETS capacity |Free ETS capacity]] [[Free ETS capacity]], ''≈E'' = ''E-L''
:::::::: [[Image:ExR.jpg|60 px|link=Excess E-R capacity|Excess ''E-R'' capacity]] [[Excess E-R capacity|Excess ''E-R'' capacity]], ''ExR'' = ''E-R''
:: [[Image:L.jpg|link=LEAK respiration|LEAK]] [[LEAK respiration]], ''L'' = ''L´''-ROX
:: [[Image:ROX.jpg|link=Residual oxygen consumption|ROX]] [[Residual oxygen consumption]], ROX (subtracted from apparent fluxes (''R´, E´, L´'')


 
{{MitoPedia concepts
=== Respiratory coupling control ratios in intact cells ===
|mitopedia concept=MiP concept, Ergodynamics
::[[Image:L over R.jpg|50 px|link=L/R coupling control ratio |''L/R'' coupling control ratio]] [[L/R coupling control ratio |''L/R'' coupling control ratio]], ''L/R''
}}
:: [[Image:L over E.jpg|50 px|link=LEAK control ratio |LEAK control ratio]] [[LEAK control ratio]], ''L/E''
{{MitoPedia methods
:: [[Image:R over E.jpg|50 px|link=ROUTINE control ratio |ROUTINE control ratio]] [[ROUTINE control ratio]], ''R/E''
|mitopedia method=Respirometry
 
}}
 
{{Labeling
=== Respiratory coupling control factors in intact cells ===
|topics=Coupling efficiency;uncoupling
:: [[Image:j--R.jpg|50 px|link=ROUTINE coupling efficiency |ROUTINE coupling efficiency]] [[ROUTINE coupling efficiency]]: ''j<sub>≈R</sub>'' = ''≈R/R'' =(''R-L'')/''R'' = 1-''L/R''
|couplingstates=LEAK, OXPHOS, ET
:: [[Image:j--E.jpg|50 px|link=ETS coupling efficiency |ETS coupling efficiency]] [[ETS coupling efficiency]], ''E-L'' control factor: ''j<sub>≈E</sub>'' = ''≈E/E'' = (''E-L'')/''E'' = 1-''L/E''
|pathways=N, S, NS, ROX
:: [[Image:jExR.jpg|50 px|link=Excess E-R capacity factor |Excess ''E-R'' capacity factor]] [[Excess E-R capacity factor |Excess ''E-R'' capacity factor]], ''E-R'' coupling control factor: ''j<sub>ExR</sub>'' = (''E-R'')/''E'' = 1-''R/E''
|instruments=Theory
:: [[Image:NetR over E.jpg|60 px|link=NetROUTINE control ratio |netROUTINE control ratio]] [[netROUTINE control ratio]], ''≈R/E'' control ratio: ''≈R/E'' = (''R-L'')/''E''
}}
 
 
[[File:EPL-free and excess.jpg|right|400px|thumb|[[Gnaiger 2014 MitoPathways |The Blue Book 2014]]: Fig. 2.4.]]
== Cell ergometry: permeabilized cells ==
=== Respiratory coupling states in mt-preparations ===
:: [[Image:P.jpg|link=OXPHOS capacity|OXPHOS]] [[OXPHOS capacity]], ''P'' = ''P´''-ROX
:::::::: [[Image:P-L.jpg|50 px|link=Free OXPHOS capacity |Free OXPHOS capacity]] [[Free OXPHOS capacity]], ''≈P'' = ''P-L''
:: [[Image:E.jpg|link=ETS capacity|ETS]] [[ETS capacity]], ''E'' = ''E´''-ROX
:::::::: [[Image:E-L.jpg|50 px|link=Free ETS capacity |Free ETS capacity]] [[Free ETS capacity]], ''≈E'' = ''E-L''
:::::::: [[Image:ExP.jpg|60 px|link=Excess E-P capacity |Excess ''E-P'' capacity]] [[Excess E-P capacity |Excess ''E-P'' capacity]], ''ExP'' = ''E-P''
::[[Image:L.jpg|link=LEAK respiration|LEAK]] [[LEAK respiration]], ''L'' = ''L´''-ROX
:: [[Image:ROX.jpg|link=Residual oxygen consumption|ROX]] [[Residual oxygen consumption]], ROX (subtracted from ''P´, E´, L´'')
 
 
=== Respiratory coupling control ratios in mt-preparations ===
::[[Image:L over P.jpg|50 px|link=L/P coupling control ratio |''L/P'' coupling control ratio]] [[L/P coupling control ratio |''L/P'' coupling control ratio]]: ''L/P''
:: [[Image:L over E.jpg|50 px|link=LEAK control ratio |LEAK control ratio]] [[LEAK control ratio]], ''L/E''
:: [[Image:P over E.jpg|50 px|link=OXPHOS control ratio |OXPHOS control ratio]] [[OXPHOS control ratio]], ''P/E''
 
 
=== Respiratory coupling control factors in mt-preparations ===
:: [[Image:j--P.jpg|50 px|link=OXPHOS coupling efficiency |OXPHOS coupling efficiency]] [[OXPHOS coupling efficiency]], (''P-L'' or ''≈P'' control factor): ''j<sub>≈P</sub>'' = ''≈P/P'' = (''P-L'')/''P'' = 1-''L/P''
:: [[Image:j--E.jpg|50 px|link=ETS coupling efficiency |ETS coupling efficiency]] [[ETS coupling efficiency]], ''E-L'' control factor: ''j<sub>≈E</sub>'' = ''≈E/E'' = (''E-L'')/''E'' = 1-''L/E''
:: [[Image:jExP.jpg|50 px|link=Excess E-P capacity factor |Excess ''E-P'' capacity factor]] [[Excess E-P capacity factor |Excess ''E-P'' capacity factor]], ''E-P'' coupling control factor: ''j<sub>ExP</sub>'' = (''E-P'')/''E'' = 1-''P/E''
:: [[Image:NetP over E.jpg|60 px|link=NetOXPHOS control ratio |netOXPHOS control ratio]] [[netOXPHOS control ratio]], ''≈P/E'' control ratio: ''≈P/E'' = (''P-L'')/''E''

Latest revision as of 01:26, 31 December 2020

{{MitoPedia |description=Biochemical cell ergometry aims at measurement of JO2max (compare VO2max or VO2peak in exercise ergometry of humans and animals) of cell respiration linked to phosphorylation of ADP to ATP. The corresponding OXPHOS capacity is based on saturating concentrations of ADP, [ADP], and inorganic phosphate [Pi] available to the mitochondria. This is metabolically opposite to uncoupling of respiration, which yields ET capacity. The OXPHOS state can be established experimentally by selective permeabilization of cell membranes with maintenance of intact mitochondria, titrations of ADP and Pi to evaluate kinetically saturating conditions, and establishing fuel substrate combinations which reconstitute physiological TCA cycle function. Uncoupler titrations are applied to determine the apparent ET-pathway excess over OXPHOS capacity ([[E-P control efficiency |E-P control efficiency) and to calculate the P-L control efficiency jP-L and E-L coupling efficiency jE-L. These normalized flux ratios are the basis to calculate the ergometric or ergodynamic efficiency, ε = j · f, where f is the normalized force ratio.

» MiPNet article |info=Gnaiger 2020 BEC MitoPathways, Oxygen flux }}

Cell ergometry and OXPHOS

Publications in the MiPMap
Gnaiger E (2015) Cell ergometry and OXPHOS. Mitochondr Physiol Network 2015-01-18.


Oroboros (2015) MiPNet

Abstract: Spiroergometry on the organismic level is compared to cell ergometry as OXPHOS analysis on the cellular level.


O2k-Network Lab: AT Innsbruck Gnaiger E


Cell ergometry.pdf

Spiroergometry

VO2max or VO2peak in cycle or treadmill spiroergometry is expressed in units of [mL O2·min-1·kg-1] body mass. 1 mL oxygen at STPD is equivalent to 22.392 mmol O2. Therefore, multiply by 1000/(22.392·60)=0.744 to convert VO2max to JO2max expressed in SI units [nmol·s-1·g-1]:
1 mL O2·min-1·kg-1 = 0.744 µmol·s-1·kg-1
VO2max (JO2max) typically declines from 70 to 25 mL O2·min-1·kg-1 (50 to 20 µmol·s-1·kg-1) in the range of healthy trained to obese untrained humans.

Keywords

  • Expand Bioblast links to Cell ergometry


Questions.jpg


Click to expand or collaps
Bioblast links: Coupling control - >>>>>>> - Click on [Expand] or [Collapse] - >>>>>>>

1. Mitochondrial and cellular respiratory rates in coupling-control states

OXPHOS-coupled energy cycles. Source: The Blue Book
» Baseline state
Respiratory rate Defining relations Icon
OXPHOS capacity P = -Rox P.jpg mt-preparations
ROUTINE respiration R = -Rox R.jpg living cells
ET capacity E = -Rox E.jpg » Level flow
» Noncoupled respiration - Uncoupler
LEAK respiration L = -Rox L.jpg » Static head
» LEAK state with ATP
» LEAK state with oligomycin
» LEAK state without adenylates
Residual oxygen consumption Rox L = -Rox ROX.jpg
  • Chance and Williams nomenclature: respiratory states
» State 1 —» State 2 —» State 3 —» State 4 —» State 5

2. Flux control ratios related to coupling in mt-preparations and living cells

» Flux control ratio
» Coupling-control ratio
» Coupling-control protocol
FCR Definition Icon
L/P coupling-control ratio L/P L/P coupling-control ratio » Respiratory acceptor control ratio, RCR = P/L
L/R coupling-control ratio L/R L/R coupling-control ratio
L/E coupling-control ratio L/E L/E coupling-control ratio » Uncoupling-control ratio, UCR = E/L (ambiguous)
P/E control ratio P/E P/E control ratio
R/E control ratio R/E R/E control ratio » Uncoupling-control ratio, UCR = E/L
net P/E control ratio (P-L)/E net P/E control ratio
net R/E control ratio (R-L)/E net R/E control ratio

3. Net, excess, and reserve capacities of respiration

Respiratory net rate Definition Icon
P-L net OXPHOS capacity P-L P-L net OXPHOS capacity
R-L net ROUTINE capacity R-L R-L net ROUTINE capacity
E-L net ET capacity E-L E-L net ET capacity
E-P excess capacity E-P E-P excess capacity
E-R reserve capacity E-R E-R reserve capacity

4. Flux control efficiencies related to coupling-control ratios

» Flux control efficiency jZ-Y
» Background state
» Reference state
» Metabolic control variable
Coupling-control efficiency Definition Icon Canonical term
P-L control efficiency jP-L = (P-L)/P = 1-L/P P-L control efficiency P-L OXPHOS-flux control efficiency
R-L control efficiency jR-L = (R-L)/R = 1-L/R R-L control efficiency R-L ROUTINE-flux control efficiency
E-L coupling efficiency jE-L = (E-L)/E = 1-L/E E-L coupling efficiency E-L ET-coupling efficiency » Biochemical coupling efficiency
E-P control efficiency jE-P = (E-P)/E = 1-P/E E-P control efficiency E-P ET-excess flux control efficiency
E-R control efficiency jE-R = (E-R)/E = 1-R/E E-R control efficiency E-R ET-reserve flux control efficiency

5. General

» Basal respiration
» Cell ergometry
» Dyscoupled respiration
» Dyscoupling
» Electron leak
» Electron-transfer-pathway state
» Hyphenation
» Oxidative phosphorylation
» Oxygen flow
» Oxygen flux
» Permeabilized cells
» Phosphorylation system
» Proton leak
» Proton slip
» Respiratory state
» Uncoupling



MitoPedia concepts: MiP concept, Ergodynamics 


MitoPedia methods: Respirometry 


Labels:




Regulation: Coupling efficiency;uncoupling  Coupling state: LEAK, OXPHOS, ET  Pathway: N, S, NS, ROX  HRR: Theory