Cookies help us deliver our services. By using our services, you agree to our use of cookies. More information

Difference between revisions of "MiPNet12.23 FiberRespiration"

From Bioblast
Β 
(39 intermediate revisions by 9 users not shown)
Line 1: Line 1:
{{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. [[Media:MiPNet12.23 FibreRespiration.pdf|Β»Bioblast pdfΒ«]]
Β 
|info=[http://www.bioblast.at/index.php/File:MiPNet12.23_FibreRespiration.pdf Versions]
|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.
|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 
:»  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 [http://www.oroboros.at/?datlab DatLab 4 software] is shown for on-line data analysis [[MiPNet12.09]] The following guideline describes the experimental protocol and includes a short discussion of results. The experiments was carried out by participants of an O2k-Course on HRR in December 2007 [http://www.oroboros.at/index.php?id=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]]
Figure 1. Oxygen concentration ([ΞΌM] blue line) and oxygen flux per mg wet weight of muscle ([pmolβˆ™s-1βˆ™mg-1] red lines) in O2k chamber B, in permeabilized fibres from horse skeletal muscle with the standard titration protocol.
Β 
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.
Β 
Β 
#<span style="color:red"> '''LEAK state, ''L'':'''</span> Non-phosphorylating resting state with substrates for Complex I (glutamate+malate; GMN; without adenalytes, N).
#<span style="color:green">'''OXPHOS capacity with Complex I substrates, ''P'':'''</span> Respiratory capacity in the active coupled state (with ADP, GMD).
#<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).
#<span style="color:green">'''OXPHOS capacity with Complex I&II substrates, ''P'':'''</span> Respiratory stimulation by convergent electron flow from Complexes I&II at the Q-junction, in the coupled state after further addition of succinate, as an estimate of OXPHOS capacity (Gnaiger 2009).
#<span style="color:blue">'''Electron transfer system (ETS) capacity with CI&II substrates, ''E'':'''</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.
#<span style="color:blue">'''ETS capacity with CII substrate, ''E'':'''</span> ETS capacity with succinate, after blocking Complex I with rotenone.
#'''Residual oxygen consumption (ROX)''' due to oxidative side reactions in the permeabilized fibres, estimated after addition of Antimycin A (Inhibitor of Complex III).
:>> Product: [http://www.oroboros.at/?oxygraph OROBOROS Oxygraph-2k], [[OROBOROS O2k-Catalogue | O2k-Catalogue]]
|mipnetlab=AT_Innsbruck_OROBOROS, BE_Liege_Votion DM
}}
{{Labeling
|area=Respiration, Instruments;methods
|organism=Horse
|tissues=Skeletal muscle
|preparations=Permeabilized tissue
|topics=Coupling efficiency;uncoupling, Cyt c
|couplingstates=LEAK, OXPHOS, ETS
|substratestates=CI, CII, CI&II
|instruments=Oxygraph-2k, O2k-Protocol
|additional=O2k-Demo, O2k-Core
}}
}}
== Further information ==
* 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]]
:>> '''[http://www.oroboros.at/?protocols_muscle_horse Excel Templates, DatLab Templates and DatLab-Demo Files]'''

Latest revision as of 10:34, 27 November 2019

                



MiPNet12.23 FiberRespiration

Publications in the MiPMap
O2k-Protocols contents
Mitochondrial respiration in permeabilized fibers: Needle biopsies from horse skeletal muscle.

Β» Bioblast pdf Β»Versions

Oroboros (2019-01-10) Mitochondr Physiol Network

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» O2k-Protocols: SUIT-011
Β» Product: Oroboros O2k, O2k-Catalogue


β€’ O2k-Network Lab: AT_Innsbruck_Oroboros, BE_Liege_Votion DM