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.|
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 (~ 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 2–3 h of cold storage.
• Keywords: High-resolution respirometry, Human skeletal muscle, Homogenate, Mitochondria, Needle biopsy
• O2k-Network Lab: CZ Prague Krajcova A
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