MiPNet12.23 FiberRespiration

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Mitochondrial respiration in permeabilized fibers: Needle biopsies from horse skeletal muscle.

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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. Methodological and conceptual features of High-Resolution Respirometry (HRR) are illustrated in an experiment with permeabilized fibers in the Oroboros O2k. The experiment demonstrates manual titrations applied to study respiratory capacity in permeabilized fibers. Application of the DatLab software is shown for real-time data analysis (compare MiPNet12.09). The following guideline describes the SUIT-011 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 (IOC44; Schroecken, Austria).

» Product: Oroboros O2k, O2k-Catalogue


O2k-Network Lab: AT_Innsbruck_Oroboros, BE_Liege_Votion DM


Labels: MiParea: Respiration, Instruments;methods 


Organism: Horse  Tissue;cell: Skeletal muscle  Preparation: Permeabilized tissue 

Regulation: Coupling efficiency;uncoupling, Cyt c  Coupling state: LEAK, OXPHOS, ET  Pathway: N, S, NS  HRR: Oxygraph-2k, O2k-Protocol 

O2k-Demo, O2k-Core, SUIT-011, SUIT-011 O2 pfi D024 

SUIT protocol

1GM;2D;2c;3S;4U;5Rot;6Ama.png

SUIT states: 1-2GM 3-4GMS 5S 6ROX

Step Respiratory state Pathway control ET-Complex entry into Q-junction
1GM GML N CI
2D GMP N CI
2c GMP N CI
3S GMSP NS CI&II
4U GMSE NS CI&II
5Rot SE S CII
6Ama ROX
MiPNet12.23.jpg

Figure 1. Oxygen concentration ([μM] blue line) and oxygen flux per mg wet weight of muscle ([pmol∙s-1∙mg-1] red lines) in O2k chamber B, in permeabilized fibers from horse skeletal muscle with the standard titration protocol.

A substrate-uncoupler-inhibitor titration (SUIT) protocol (Fig. 1) was developed for respiratory studies of permeabilized muscle fibers. The protocol aims at

Reference

  • 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. - »Bioblast link«