An optimized protocol for coupling oxygen consumption rates with β-oxidation in isolated mitochondria from mouse soleus
Summary: Depending on metabolic requirements, skeletal muscle mitochondria integrate O2 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 meas...
| Main Authors: | , |
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| Format: | Article |
| Language: | English |
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Elsevier
2021-09-01
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| Series: | STAR Protocols |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166721004421 |
| _version_ | 1819132539426570240 |
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| author | Cristina Sánchez-González Laura Formentini |
| author_facet | Cristina Sánchez-González Laura Formentini |
| author_sort | Cristina Sánchez-González |
| collection | DOAJ |
| description | Summary: Depending on metabolic requirements, skeletal muscle mitochondria integrate O2 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 O2 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). |
| first_indexed | 2024-12-22T09:33:00Z |
| format | Article |
| id | doaj.art-ac5b86f1ba694f91ad9b76b51c50139b |
| institution | Directory Open Access Journal |
| issn | 2666-1667 |
| language | English |
| last_indexed | 2024-12-22T09:33:00Z |
| publishDate | 2021-09-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj.art-ac5b86f1ba694f91ad9b76b51c50139b2022-12-21T18:30:53ZengElsevierSTAR Protocols2666-16672021-09-0123100735An optimized protocol for coupling oxygen consumption rates with β-oxidation in isolated mitochondria from mouse soleusCristina Sánchez-González0Laura Formentini1Departamento de Biología Molecular, Centro de Biología Molecular “Severo Ochoa” (CBMSO), Universidad Autónoma de Madrid, c/ Nicolas Cabrera 1, 28049, Madrid, Spain; Corresponding authorDepartamento de Biología Molecular, Centro de Biología Molecular “Severo Ochoa” (CBMSO), Universidad Autónoma de Madrid, c/ Nicolas Cabrera 1, 28049, Madrid, Spain; Centro de Investigación Biomédica en red de Enfermedades Raras (CIBERER), ISCIII, Madrid, Spain; Instituto de Investigación Hospital 12 de Octubre, i+12, Universidad Autónoma de Madrid, Madrid, Spain; Corresponding authorSummary: Depending on metabolic requirements, skeletal muscle mitochondria integrate O2 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 O2 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).http://www.sciencedirect.com/science/article/pii/S2666166721004421cell biologycell isolationcell membranecell separation/fractionationmetabolism |
| spellingShingle | Cristina Sánchez-González Laura Formentini An optimized protocol for coupling oxygen consumption rates with β-oxidation in isolated mitochondria from mouse soleus STAR Protocols cell biology cell isolation cell membrane cell separation/fractionation metabolism |
| title | An optimized protocol for coupling oxygen consumption rates with β-oxidation in isolated mitochondria from mouse soleus |
| title_full | An optimized protocol for coupling oxygen consumption rates with β-oxidation in isolated mitochondria from mouse soleus |
| title_fullStr | An optimized protocol for coupling oxygen consumption rates with β-oxidation in isolated mitochondria from mouse soleus |
| title_full_unstemmed | An optimized protocol for coupling oxygen consumption rates with β-oxidation in isolated mitochondria from mouse soleus |
| title_short | An optimized protocol for coupling oxygen consumption rates with β-oxidation in isolated mitochondria from mouse soleus |
| title_sort | optimized protocol for coupling oxygen consumption rates with β oxidation in isolated mitochondria from mouse soleus |
| topic | cell biology cell isolation cell membrane cell separation/fractionation metabolism |
| url | http://www.sciencedirect.com/science/article/pii/S2666166721004421 |
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