An optimized protocol for coupling oxygen consumption rates with β-oxidation in isolated mitochondria from mouse soleus .

Depending on metabolic requirements, skeletal muscle mitochondria integrate O ₂ consumption and ATP production with lipid, glucose, or amino acid metabolism. Free fatty acids (FFAs) are the main source of energy during rest and mild-intensity exercise. We present a detailed protocol for measuring FFA-β-oxidation coupled with O ₂ respiration by a Clark-type electrode in isolated mitochondria from mouse soleus oxidative muscle. We optimized the procedure, including buffer composition, protease treatment, and quantifiable parameters (P/O, Phosphate/Oxygen Ratio; OCR, Oxygen Consumption Rate; RCR,Respiration Control Rate; OSR, Oligomycin Sensitive Respiration). For complete details on the use and execution of this protocol, please refer to Sanchez-Gonzalez et al. (2020).
Competing Interests: The authors declare no competing interests.
(© 2021 The Author(s).)