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Assessing the function of mitochondria in cytosolic context in human skeletal muscle: adopting high-resolution respirometry to homogenate of needle biopsy tissue samples
J. Ziak, A. Krajcova, K. Jiroutkova, V. Nemcova, V. Dzupa, F. Duska,
Jazyk angličtina Země Nizozemsko
Typ dokumentu časopisecké články, práce podpořená grantem
Grantová podpora
NT12319
MZ0
CEP - Centrální evidence projektů
- MeSH
- buněčné dýchání * MeSH
- buněčné extrakty MeSH
- cytologické techniky metody MeSH
- dospělí MeSH
- jehlová biopsie MeSH
- kosterní svaly fyziologie MeSH
- lidé středního věku MeSH
- lidé MeSH
- mitochondriální nemoci diagnóza MeSH
- mitochondrie fyziologie MeSH
- odběr biologického vzorku metody MeSH
- senioři nad 80 let MeSH
- senioři MeSH
- Check Tag
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- senioři nad 80 let MeSH
- senioři MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Using skeletal muscle homogenates for respirometry has many advantages, but the main challenge is avoiding the damage to outer mitochondrial membrane (OMM) and complex I. By optimising the amount of muscle and careful titration of substrates and inhibitors we developed a new protocol and compared it to isolated mitochondria. We found acceptable damage to OMM (~10-15% increment of oxygen flux after addition of cytochrome c) and to complex I (~70% of electron flux). Homogenate retained ~90% of phosphorylation capacity of isolated mitochondria. The use of fresh homogenate was crucial as mitochondrial function declined rapidly after 2-3h of cold storage.
Department of Molecular Biology 3rd Faculty of Medicine Charles University Prague Czech Republic
Department of Orthopaedic Surgery Kralovske Vinohrady University Hospital Prague Czech Republic
Laboratory of Bioenergetics 3rd Faculty of Medicine Charles University Prague Czech Republic
Citace poskytuje Crossref.org
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