In situ immunofluorescent staining of autophagy in muscle stem cells

Francesco Castagnetti, Elisabetta Fiacco, Carol Imbriano, Lucia Latella

Research output: Contribution to journalArticle

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.

Original languageEnglish
Article numbere55908
JournalJournal of Visualized Experiments
Volume2017
Issue number124
DOIs
Publication statusPublished - Jun 12 2017

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Keywords

  • Autophagy
  • Cellular Biology
  • Issue 124
  • LC3
  • Muscle regeneration
  • Muscle stem cells (satellites)
  • Muscle tissue sections
  • MYOD

ASJC Scopus subject areas

  • Neuroscience(all)
  • Chemical Engineering(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Immunology and Microbiology(all)

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