Ziak 2015 Mitochondrion
Ziak J, Krajcova A, Jiroutkova K, Nemcova V, Dzupa V, Duska F (2015) Assessing the function of mitochondria in cytosolic context in human skeletal muscle: Adopting high-resolution respirometry to homogenate of needle biopsy tissue samples. Mitochondrion 21:106-12. |
Ziak J, Krajcova A, Jiroutkova K, Nemcova V, Dzupa V, Duska F (2015) Mitochondrion
Abstract: 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 (~ 10β15% 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 2β3 h of cold storage. β’ Keywords: High-resolution respirometry, Human skeletal muscle, Homogenate, Mitochondria, Needle biopsy
Labels: MiParea: Respiration, mt-Membrane
Organism: Human
Tissue;cell: Skeletal muscle
Preparation: Homogenate, Isolated mitochondria
Coupling state: LEAK, OXPHOS, ET
Pathway: N, S, NS, ROX
HRR: Oxygraph-2k