Assessing autophagy in muscle stem cells

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• dc.contributor.author Campanario, Silvia
• dc.contributor.author Ramírez Pardo, Ignacio
• dc.contributor.author Hong, Xiaotong
• dc.contributor.author Isern, Joan
• dc.contributor.author Muñoz Cánoves, Pura, 1962-
• dc.date.accessioned 2021-03-08T06:54:25Z
• dc.date.available 2021-03-08T06:54:25Z
• dc.date.issued 2021
• dc.description.abstract The skeletal muscle tissue in the adult is relatively stable under normal conditions but retains a striking ability to regenerate by its resident stem cells (satellite cells). Satellite cells exist in a quiescent (G0) state; however, in response to an injury, they reenter the cell cycle and start proliferating to provide sufficient progeny to form new myofibers or undergo self-renewal and returning to quiescence. Maintenance of satellite cell quiescence and entry of satellite cells into the activation state requires autophagy, a fundamental degradative and recycling process that preserves cellular proteostasis. With aging, satellite cell regenerative capacity declines, correlating with loss of autophagy. Enhancing autophagy in aged satellite cells restores their regenerative functions, underscoring this proteostatic activity's relevance for tissue regeneration. Here we describe two strategies for assessing autophagic activity in satellite cells from GFP-LC3 reporter mice, which allows direct autophagosome labeling, or from non-transgenic (wild-type) mice, where autophagosomes can be immunostained. Treatment of GFP-LC3 or WT satellite cells with compounds that interfere with autophagosome-lysosome fusion enables measurement of autophagic activity by flow cytometry and immunofluorescence. Thus, the methods presented permit a relatively rapid assessment of autophagy in stem cells from skeletal muscle in homeostasis and in different pathological scenarios such as regeneration, aging or disease.
• dc.description.sponsorship Work in the authors’ laboratory has been supported by MINECO-Spain (RTI2018-096068), ERC-2016-AdG-741966, LaCaixa-HEALTH-HR17-00040, MDA, Fundació LaMarató/TV3, MDA, UPGRADE-H2020-825825, AFM and DPP-Spain, as well as María-de-Maeztu-Program for Units of Excellence to UPF (MDM-2014-0370), Severo-Ochoa-Program for Centers of Excellence to CNIC (SEV-2015-0505). SC was supported by a Predoctoral Fellowship from Ayudas para la formación y contratación de personal investigador novel (FI) – AGAUR (Barcelona, Spain), IR-P was supported by a Predoctoral Fellowship from Programa de Formación de Personal Investigador (Spain), and XH was supported by a Severo-Ochoa Pre-doctoral Fellowship (Spain).
• dc.format.mimetype application/pdf
• dc.identifier.citation Campanario S, Ramírez-Pardo I, Hong X, Isern J, Muñoz-Cánoves P. Assessing autophagy in muscle stem cells. Front Cell Dev Biol. 2021; 8:620409. DOI: 10.3389/fcell.2020.620409
• dc.identifier.doi http://dx.doi.org/10.3389/fcell.2020.620409
• dc.identifier.issn 2296-634X
• dc.identifier.uri http://hdl.handle.net/10230/46683
• dc.language.iso eng
• dc.publisher Frontiers
• dc.relation.ispartof Front Cell Dev Biol. 2021; 8:620409
• dc.relation.projectID info:eu-repo/grantAgreement/EC/H2020/741966
• dc.relation.projectID info:eu-repo/grantAgreement/EC/H2020/825825
• dc.rights © 2021 Campanario, Ramírez-Pardo, Hong, Isern and Muñoz-Cánoves. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
• dc.rights.accessRights info:eu-repo/semantics/openAccess
• dc.rights.uri http://creativecommons.org/licenses/by/4.0/
• dc.subject.keyword Autophagy
• dc.subject.keyword Flow cytometry
• dc.subject.keyword Immunofluorescence
• dc.subject.keyword Quiescence
• dc.subject.keyword Regeneration
• dc.subject.keyword Satellite cell
• dc.subject.keyword Skeletal muscle
• dc.subject.keyword Stem cell
• dc.title Assessing autophagy in muscle stem cells
• dc.type info:eu-repo/semantics/article
• dc.type.version info:eu-repo/semantics/publishedVersion