Burtscher 2014 Abstract MiP2014
|Region-specific differences in Complex I- and Complex II-linked respiration in the mouse brain.|
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-pathway capacity (ET-pathway) 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-pathway (ET-pathway) capacity in a single run from as little as 2 mg tissue applying the Oroboros high-resolution respirometry system . 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-linked ET capacity and CIV activity expressed as oxygen consumption per tissue mass or as CIV/CI&IIE flux control ratios. The P/E coupling control ratio (CI&II), an index of the limitation of OXPHOS capacity by the phorphorylation system, was significantly different between motorcortex and hippocampus.
The established protocol allows detailed analysis of mitochondrial function from small amounts of specific tissues. It thus enables comparison of different brain regions implicated in neurodegenerative diseases of the healthy mouse and disease models while leaving sufficient amounts of sample for additional analysis of the tissues.
Labels: MiParea: Respiration
Organism: Mouse Tissue;cell: Nervous system Preparation: Permeabilized tissue
Coupling state: OXPHOS, ET Pathway: N, S, CIV, NS HRR: Oxygraph-2k Event: A4, Oral MiP2014
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).
- Pesta D, Gnaiger E (2012) High-resolution respirometry. OXPHOS protocols for human cells and permeabilized fibres from small biopsies of human muscle. Methods Mol Biol 810:25-58.