ArticleActa physiologica (Oxford, England)2019
Isolation of mitochondrial subpopulations from skeletal muscle: Optimizing recovery and preserving integrity.
Article in Acta physiologica (Oxford, England), 2019. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 22 papers.
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Who cites it
22 citing papers in PubMed.
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- Mitochondrial Metabolomics in Cancer: Mass Spectrometry-Based Approaches for Metabolic Rewiring Analysis and Therapeutic Discovery.Metabolites · 2025Review
- Hypotonic Swelling Method for the Isolation of Pure Mitochondria From Primary Human Skeletal Myoblasts for Proteomic Studies.Journal of cellular and molecular medicine · 2025Article
- The Worsening of Myocardial Ischemia-Reperfusion Injury in Uremic Cardiomyopathy is Further Aggravated by PMCardiovascular toxicology · 2024Article
- Mitochondrial mechanisms in the pathogenesis of chronic inflammatory musculoskeletal disorders.Cell & bioscience · 2024Review
- Impact of capillary and sarcolemmal proximity on mitochondrial structure and energetic function in skeletal muscle.The Journal of physiology · 2024Article
- A pilot study of mitochondrial response to an in vivo prosthetic joint Staphylococcus aureus infection model.Journal of orthopaedic research : official publication of the Orthopaedic Research Society · 2024Article
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- Mitochondria isolated from lipid droplets of white adipose tissue reveal functional differences based on lipid droplet size.Life science alliance · 2024Article
- The Predictive Value of Graft Viability and Bioenergetics Testing Towards the Outcome in Liver Transplantation.Transplant international : official journal of the European Society for Organ Transplantation · 2024Article
- Cholic and deoxycholic acids induce mitochondrial dysfunction, impaired biogenesis and autophagic flux in skeletal muscle cells.Biological research · 2023Article
- Isolation of Mitochondria from Mouse Tissues for Functional Analysis.Methods in molecular biology (Clifton, N.J.) · 2023Article
- Blood volume versus deoxygenated NIRS signal: computational analysis of the effects muscle OJournal of applied physiology (Bethesda, Md. : 1985) · 2021Article
- Subcellular Specialization of Mitochondrial Form and Function in Skeletal Muscle Cells.Frontiers in cell and developmental biology · 2021Review
- Bioenergetic functions in subpopulations of heart mitochondria are preserved in a non-obese type 2 diabetes rat model (Goto-Kakizaki).Scientific reports · 2020Article
- Integrated approach for data acquisition, visualization and processing of analog polarographic systems for bioenergetics studies.Analytical biochemistry · 2020Article
- Rapid Single-Step Affinity Purification of HA-Tagged Plant Mitochondria.Plant physiology · 2020Article
- Isolation of mitochondria from cells and tissues.Methods in cell biology · 2020Article
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6 authors.
Funding
Abstract
aimThe subsarcolemmal (SSM) and interfibrillar (IFM) mitochondria in skeletal muscle appear to have distinct biochemical properties affecting metabolism in health and disease. The isolation of mitochondrial subpopulations has been a long-time challenge while the presence of a continuous mitochondrial reticulum challenges the view of distinctive SSM and IFM bioenergetics. Here, a comprehensive approach is developed to identify the best conditions to separate mitochondrial fractions.
methodsThe main modifications to the protocol to isolate SSM and IFM from rat skeletal muscle were: (a) decreased dispase content and homogenization speed; (b) trypsin treatment of SSM fractions; (c) recentrifugation of mitochondrial fractions at low speed to remove subcellular components. To identify the conditions preserving mitochondrial function, integrity, and maximizing their recovery, microscopy (light and electron) were used to monitor effectiveness and efficiency in separating mitochondrial subpopulations while respiratory and enzyme activities were employed to evaluate function, recovery, and integrity.
resultsWith the modifications described, the total mitochondrial yield increased with a recovery of 80% of mitochondria contained in the original skeletal muscle sample. The difference between SSM and IFM oxidative capacity (10%) with complex-I substrate was significant only with a saturated ADP concentration. The inner and outer membrane damage for both subpopulations was <1% and 8%, respectively, while the respiratory control ratio was 16.
conclusionUsing a multidisciplinary approach, conditions were identified to maximize SSM and IFM recovery while preserving mitochondrial integrity, biochemistry, and morphology. High quality and recovery of mitochondrial subpopulations allow to study the relationship between these organelles and disease.
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