In Situ Immunofluorescent Staining of Autophagy in Muscle Stem Cells

Handle URI:
http://hdl.handle.net/10754/625608
Title:
In Situ Immunofluorescent Staining of Autophagy in Muscle Stem Cells
Authors:
Castagnetti, Francesco; Fiacco, Elisabetta; Imbriano, Carol; Latella, Lucia
Abstract:
Increasing evidence points to autophagy as a crucial regulatory process to preserve tissue homeostasis. It is known that autophagy is involved in skeletal muscle development and regeneration, and the autophagic process has been described in several muscular pathologies and agerelated muscle disorders. A recently described block of the autophagic process that correlates with the functional exhaustion of satellite cells during muscle repair supports the notion that active autophagy is coupled with productive muscle regeneration. These data uncover the crucial role of autophagy in satellite cell activation during muscle regeneration in both normal and pathological conditions, such as muscular dystrophies. Here, we provide a protocol to monitor the autophagic process in the adult Muscle Stem Cell (MuSC) compartment during muscle regenerative conditions. This protocol describes the setup methodology to perform in situ immunofluorescence imaging of LC3, an autophagy marker, and MyoD, a myogenic lineage marker, in muscle tissue sections from control and injured mice. The methodology reported allows for monitoring the autophagic process in one specific cell compartment, the MuSC compartment, which plays a central role in orchestrating muscle regeneration.
KAUST Department:
Biological and Environmental Sciences and Engineering (BESE) Division
Citation:
Castagnetti F, Fiacco E, Imbriano C, Latella L (2017) <em>In Situ</em> Immunofluorescent Staining of Autophagy in Muscle Stem Cells. Journal of Visualized Experiments. Available: http://dx.doi.org/10.3791/55908.
Publisher:
MyJove Corporation
Journal:
Journal of Visualized Experiments
Issue Date:
13-Jun-2017
DOI:
10.3791/55908
Type:
Article
ISSN:
1940-087X
Sponsors:
This work was supported by NIAMS AR064873, Epigen Project PB. P01.001.019/Progetto Bandiera Epigenomica IFT to L.L.
Additional Links:
https://www.jove.com/video/55908/in-situ-immunofluorescent-staining-of-autophagy-in-muscle-stem-cells
Appears in Collections:
Articles; Biological and Environmental Sciences and Engineering (BESE) Division

Full metadata record

DC FieldValue Language
dc.contributor.authorCastagnetti, Francescoen
dc.contributor.authorFiacco, Elisabettaen
dc.contributor.authorImbriano, Carolen
dc.contributor.authorLatella, Luciaen
dc.date.accessioned2017-10-03T12:49:29Z-
dc.date.available2017-10-03T12:49:29Z-
dc.date.issued2017-06-13en
dc.identifier.citationCastagnetti F, Fiacco E, Imbriano C, Latella L (2017) <em>In Situ</em> Immunofluorescent Staining of Autophagy in Muscle Stem Cells. Journal of Visualized Experiments. Available: http://dx.doi.org/10.3791/55908.en
dc.identifier.issn1940-087Xen
dc.identifier.doi10.3791/55908en
dc.identifier.urihttp://hdl.handle.net/10754/625608-
dc.description.abstractIncreasing evidence points to autophagy as a crucial regulatory process to preserve tissue homeostasis. It is known that autophagy is involved in skeletal muscle development and regeneration, and the autophagic process has been described in several muscular pathologies and agerelated muscle disorders. A recently described block of the autophagic process that correlates with the functional exhaustion of satellite cells during muscle repair supports the notion that active autophagy is coupled with productive muscle regeneration. These data uncover the crucial role of autophagy in satellite cell activation during muscle regeneration in both normal and pathological conditions, such as muscular dystrophies. Here, we provide a protocol to monitor the autophagic process in the adult Muscle Stem Cell (MuSC) compartment during muscle regenerative conditions. This protocol describes the setup methodology to perform in situ immunofluorescence imaging of LC3, an autophagy marker, and MyoD, a myogenic lineage marker, in muscle tissue sections from control and injured mice. The methodology reported allows for monitoring the autophagic process in one specific cell compartment, the MuSC compartment, which plays a central role in orchestrating muscle regeneration.en
dc.description.sponsorshipThis work was supported by NIAMS AR064873, Epigen Project PB. P01.001.019/Progetto Bandiera Epigenomica IFT to L.L.en
dc.publisherMyJove Corporationen
dc.relation.urlhttps://www.jove.com/video/55908/in-situ-immunofluorescent-staining-of-autophagy-in-muscle-stem-cellsen
dc.subjectAutophagyen
dc.subjectCellular Biologyen
dc.subjectIssue 124en
dc.subjectLC3en
dc.subjectMuscle regenerationen
dc.subjectMuscle stem cells (satellites)en
dc.subjectMuscle tissue sectionsen
dc.subjectMYODen
dc.titleIn Situ Immunofluorescent Staining of Autophagy in Muscle Stem Cellsen
dc.typeArticleen
dc.contributor.departmentBiological and Environmental Sciences and Engineering (BESE) Divisionen
dc.identifier.journalJournal of Visualized Experimentsen
dc.contributor.institutionEpigenetics and Regenerative Medicine, IRCCS Fondazione Santa Lucia, , Italyen
dc.contributor.institutionDepartment of Medicine, Institute of Translational Pharmacology, Italian National Research Council, , Italyen
dc.contributor.institutionDepartment of Life Sciences, University of Modena and Reggio Emilia, , Italyen
kaust.authorFiacco, Elisabettaen
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