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Difference between revisions of "Burtscher 2014 Abstract MiP2014"

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|preparations=Permeabilized tissue
|preparations=Permeabilized tissue
|couplingstates=OXPHOS, ETS
|couplingstates=OXPHOS, ETS
|substratestates=CI, CII, CIV, CI+II
|instruments=Oxygraph-2k
|instruments=Oxygraph-2k
|additional=MiP2014
|additional=MiP2014

Revision as of 08:25, 8 August 2014

Region-specific differences in Complex I- and Complex II-linked respiration in the mouse brain.

Link:

MiP2014, Book of Abstracts Open Access

Burtscher J, Heidler J, Gnaiger E, Schwarzer C (2014)

Event: MiP2014

Mitochondrial dysfunction appears to be a common factor in neurodegenerative diseases. However, such diseases differ markedly in the nervous tissue affected. To test potential differences in mitochondrial respiratory capacity of different brain tissues under physiological or pathological conditions, we established a SUIT protocol for the analysis of oxidative phosphorylation (OXPHOS) and electron transfer system capacity (ETS) of small amounts of defined brain-tissues of mice. This protocol enables us to measure, independently, Complex I-, II- and IV-linked (CI, CII, and CIV, respectively) respiration, as well as the combined CI+II-linked OXPHOS- and electron transfer system (ETS) capacity in a single run from as little as 2 mg tissue applying the OROBOROS high-resolution respirometry system [1]. The reproducibility within one experiment (two replica from the same tissue sample) and between experiments was very high. We observed significantly higher CI-linked oxygen fluxes in the motorcortex and CII-linked respiration in the striatum, when comparing motorcortex, striatum, hippocampus and brainstem obtained from young, healthy, adult, male C57BL6/J mice. No differences were found for CI+II ETS-capacity and CIV-activity expressed as oxygen consumption per tissue mass or as CIV/CI+II flux control ratio. Motorcortex and hippocampus differed in their OXPHOS capacity when normalized to the ETS capacity. The established protocol allows detailed analysis of mitochondrial function from small amounts of specific tissues. It thus enables comparison of different brain tissues implicated in neurodegenerative diseases of the healthy mouse and disease models while leaving sufficient amounts of sample for additional analysis of the tissues.


β€’ O2k-Network Lab: AT Innsbruck Gnaiger E, AT Innsbruck OROBOROS, AT Innsbruck MitoCom


Labels: MiParea: Respiration 


Organism: Mouse  Tissue;cell: Nervous system  Preparation: Permeabilized tissue 


Coupling state: OXPHOS, ETS"ETS" is not in the list (LEAK, ROUTINE, OXPHOS, ET) of allowed values for the "Coupling states" property. 

HRR: Oxygraph-2k 

MiP2014 

Affiliation

1-Dep Pharmacol*; 2-Dep General and Transplant Surgery*; *Medical Univ Innsbruck; 3-OROBOROS Instruments; Innsbruck, Austria. - Johannes.Burtscher@i-med.ac.at

References and acknowledgements

Supported by FWF W1206-B05 (CS) and K-Regio project MitoCom Tyrol (EG).

  1. Pesta D, Gnaiger E (2012) High-resolution respirometry. OXPHOS protocols for human cells and permeabilized fibres from small biopisies of human muscle. Methods Mol Biol 810: 25-58.