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| | {{OROBOROS header page name}} |
| {{Publication | | {{Publication |
| |title=[[Image:O2k-Protocols.jpg|right|80px|link=O2k-Protocols|O2k-Protocols contents]]Mitochondrial respiration in permeabilized fibres: Needle biopsies from horse skeletal muscle. | | Β |
| |info=[[File:PDF.jpg|100px|link=http://wiki.oroboros.at/images/4/49/MiPNet12.23_FibreRespiration.pdf|Bioblast pdf]] Β | | |title=[[Image:O2k-Protocols.jpg|right|80px|link=O2k-Protocols|O2k-Protocols contents]]Mitochondrial respiration in permeabilized fibers: Needle biopsies from horse skeletal muscle. |
| |authors=OROBOROS | | |info=[[File:PDF.jpg|100px|link=http://wiki.oroboros.at/images/4/49/MiPNet12.23_FibreRespiration.pdf|Bioblast pdf]] Β»[http://www.bioblast.at/index.php/File:MiPNet12.23_FibreRespiration.pdf Versions] |
| |year=2014-11-07 | | |authors=Oroboros |
| | |year=2019-01-10 |
| |journal=Mitochondr Physiol Network | | |journal=Mitochondr Physiol Network |
| |abstract='''Lemieux H, Votion DM, Gnaiger E (2014) Mitochondrial respiration in permeabilized fibres: Needle biopsies from horse skeletal muscle. Mitochondr Physiol Network 12.23(07):1-8.''' Β»[http://www.bioblast.at/index.php/File:MiPNet12.23_FibreRespiration.pdf Versions] | | |abstract=Lemieux H, Votion DM, Doerrier C, Gnaiger E (2019) Mitochondrial respiration in permeabilized fibers: Needle biopsies from horse skeletal muscle. Mitochondr Physiol Network 12.23(08):1-4. |
| Β | | :Β» O2k-Protocols:[[SUIT-011| SUIT-011]] |
| Methodological and conceptual features of highresolution respirometry are illustrated in an experiment with permeabilized fibres in the OROBOROS O2k. The experiment demonstrates [http://bioblast.at/images/e/ef/MiPNet09.12_O2k-Titrations.pdf manual titrations] applied to study respiratory capacity in permeabilized fibres. Application of the [[DatLab]] software is shown for real-time data analysis (compare [[MiPNet12.09]]). The following guideline describes the SUIT protocol and includes a short discussion of results (see [[Votion 2012 PLoS One]]). The experiment was carried out by participants of an O2k-Workshop on HRR in December 2007 ([[MiPNet12.24 IOC44 |IOC44]]; Schroecken, Austria).
| | :Β» Product: [[Oroboros O2k]], [[Oroboros O2k-Catalogue | O2k-Catalogue]] |
| Β | | |mipnetlab=AT_Innsbruck_Oroboros, BE_Liege_Votion DM |
| [[Image:MiPNet12.23.jpg|center|100px|frame]]
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| Figure 1. Oxygen concentration ([ΞΌM] blue line) and oxygen flux per mg wet weight of muscle ([pmolβs-1βmg-1] red lines) in O2k chamber B, in permeabilized fibres from horse skeletal muscle with the standard titration protocol.
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| A multiple substrate-uncoupler-inhibitor titration protocol (Fig. 1) was developed for respiratory studies of permeabilized muscle fibres. The experiment was aimed at an assessment of a sequence of titrations, inducing stepwise defined respiratory states.
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| #<span style="color:red"> '''LEAK state with type N substrates, N''<sub>L</sub>'':'''</span> Non-phosphorylating resting state with NADH-linked (type N) substrates glutamate&malate (GM; without adenalytes; CI-linked pathway to Q).
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| #<span style="color:green">'''OXPHOS capacity with type N substrates, N''<sub>P</sub>'':'''</span> Respiratory capacity in the active coupled state (GM with ADP).
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| #<span style="color:black">'''Cytochrome ''c'' test for quality control:'''</span> Further addition of cytochrome ''c'' yields a test for integrity of the outer mitochondrial membrane (loss of cytochrome ''c'' would be indicated by a stimulation of respiration).
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| #<span style="color:green">'''OXPHOS capacity with type NS substrates (CI<small>&</small>II-linked pathway to Q), NS''<sub>P</sub>'':'''</span> Respiratory stimulation by convergent electron flow through Complexes I<small>&</small>II at the Q-junction, in the coupled state after further addition of succinate (S), as an estimate of OXPHOS capacity with reconstitution of the TCA cycle ([[Gnaiger 2009 Int J Biochem Cell Biol]]).
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| #<span style="color:blue">'''Electron transfer system (ETS) capacity with type NS substrates, NS''<sub>E</sub>'':'''</span> Uncoupling by FCCP titration (avoiding inhibition by high FCCP concentrations), as a test for limitation of OXPHOS relative to ETS capacity by the phosphorylation system.
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| #<span style="color:blue">'''ETS capacity with type S (CII) substrate, S''<sub>E</sub>'':'''</span> ETS capacity with succinate, after blocking Complex I with rotenone.
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| #'''Residual oxygen consumption (ROX)''' due to oxidative side reactions in the permeabilized fibres, estimated after addition of Antimycin A (inhibitor of Complex III).
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| :Β» Product: [[Oxygraph-2k |OROBOROS Oxygraph-2k]], [[OROBOROS O2k-Catalogue | O2k-Catalogue]] | |
| |mipnetlab=AT_Innsbruck_OROBOROS, BE_Liege_Votion DM | |
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| {{Labeling
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| |area=Respiration, Instruments;methods
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| |organism=Horse
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| |tissues=Skeletal muscle
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| |preparations=Permeabilized tissue
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| |topics=Coupling efficiency;uncoupling, Cyt c
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| |couplingstates=LEAK, OXPHOS, ETS
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| |substratestates=CI, CII, CI&II
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| |instruments=Oxygraph-2k, O2k-Protocol
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| |additional=O2k-Demo, O2k-Core
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| }} | | }} |
| == Further information ==
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| ::::* Votion DM, Gnaiger E, Lemieux H, Mouithys-Mickalad A, Serteyn D (2012) Physical fitness and mitochondrial respiratory capacity in horse skeletal muscle. PLoS One 7: e34890. - [[Votion 2012 PLoS One |Open Access]]
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| ::::Β» '''[http://www.oroboros.at/?protocols_muscle_horse Excel Templates, DatLab Templates and DatLab-Demo Files]'''
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Β»Versions
