title: Methods for Mitochondria and Mitophagy Flux Analyses in Stem Cells of Resting and Regenerating Skeletal Muscle.
abstract: Mitochondria generate most of the cell's supply of ATP as a source of energy. They are also implicated in the control of cell's growth and death. Because of these critical functions, mitochondrial fitness is key for cellular homeostasis. Often, however, mitochondria become defective following damage or stress. To prevent accumulation of damaged mitochondria, the cells clear them through mitophagy, which is defined as the selective degradation of mitochondria by autophagy (the process for degradation of long-lived proteins and damaged organelles in lysosomes). Recently, constitutive mitophagic activity has been reported in quiescent muscle stem cells (satellite cells), which sustain regeneration of skeletal muscle. In response to muscle damage, these cells activate, expand, and differentiate to repair damaged myofibers. Mitophagy was shown to be required for maintenance of satellite cells in their healthy quiescent state. Conversely, damaged mitochondria accumulated in satellite cells with aging and this was attributed to defective mitophagy. This caused increased levels of reactive oxygen species (ROS) and loss of muscle stem cell regenerative capacity at old age. In this chapter, we describe different experimental strategies to evaluate mitochondria status and mitophagy in muscle stem cells from mice. They should improve our ability to study muscle stem homeostasis in adult life, and their loss of function in aging and disease.
authors: Cell Biology Group, Department of Experimental and Health Sciences, Pompeu Fabra University (UPF), CIBER on Neurodegenerative diseases (CIBERNED), Plaça de la Mercè, 10, 08002, Barcelona, Spain.
authors: Laura
authors: L
authors: García-Prat
authors: Neurodegenerative Diseases Research Group, Vall d'Hebron Research Institute-CIBERNED, Mediterrania Building, Lab.102, Passeig de la Vall d'Hebron, 119-129, 08035, Barcelona, Spain. marta.martinez@vhir.org.
authors: Marta
authors: M
authors: Martínez-Vicente
authors: Cell Biology Group, Department of Experimental and Health Sciences, Pompeu Fabra University (UPF), CIBER on Neurodegenerative diseases (CIBERNED), Plaça de la Mercè, 10, 08002, Barcelona, Spain. pura.munoz@upf.edu.|Institució Catalana de Recerca i Estudis Avançats (ICREA), Barcelona, Spain. pura.munoz@upf.edu.
authors: Pura
authors: P
authors: Muñoz-Cánoves
mesh_terms: D001343:Autophagy; D015415:Biomarkers; D002469:Cell Separation; D016130:Immunophenotyping; D053078:Membrane Potential, Mitochondrial; D008928:Mitochondria / Q000378:metabolism*; D063306:Mitophagy; D018482:Muscle, Skeletal / Q000166:cytology* / Q000378:metabolism*; D017382:Reactive Oxygen Species / Q000378:metabolism; D012038:Regeneration; D016192:Resting Phase, Cell Cycle; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism*
chemical_list: D015415:Biomarkers; D017382:Reactive Oxygen Species
keywords: Autophagy; Mitochondria; Mitocondria membrane potential; Mitophagy; Reactive oxygen species (ROS); Satellite cell; Skeletal muscle regeneration; Stem cell