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    In Situ Immunofluorescent Staining of Autophagy in Muscle Stem Cells

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    Type
    Article
    Authors
    Castagnetti, Francesco
    Fiacco, Elisabetta
    Imbriano, Carol
    Latella, Lucia
    KAUST Department
    Biological and Environmental Sciences and Engineering (BESE) Division
    Date
    2017-06-12
    Permanent link to this record
    http://hdl.handle.net/10754/625608
    
    Metadata
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    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.
    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.
    Sponsors
    This work was supported by NIAMS AR064873, Epigen Project PB. P01.001.019/Progetto Bandiera Epigenomica IFT to L.L.
    Publisher
    MyJove Corporation
    Journal
    Journal of Visualized Experiments
    DOI
    10.3791/55908
    PubMed ID
    28654079
    Additional Links
    https://www.jove.com/video/55908/in-situ-immunofluorescent-staining-of-autophagy-in-muscle-stem-cells
    ae974a485f413a2113503eed53cd6c53
    10.3791/55908
    Scopus Count
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    Articles; Biological and Environmental Science and Engineering (BESE) Division

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