Detection of muscle stem cell-derived myonuclei in murine overloaded muscles

• Published in: STAR protocols Vol. 3; no. 2; p. 101307
• Main Authors: Kaneshige, Akihiro, Kaji, Takayuki, Saito, Hayato, Higashimoto, Tatsuyoshi, Nakamura, Ayasa, Kurosawa, Tamaki, Ikemoto-Uezumi, Madoka, Uezumi, Akiyoshi, Fukada, So-ichiro
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 17.06.2022; Elsevier
• Subjects: Animals; Cell Biology; Cell Nucleus; Mice; Microscopy; Muscle, Skeletal; Protocol; Satellite Cells, Skeletal Muscle; Stem Cells; Stem Cells; Microscopy; Cell Biology
• ISSN: 2666-1667; 2666-1667
• DOI: 10.1016/j.xpro.2022.101307

Muscle satellite cells (MuSCs) supply nuclei to existing myofibers in response to mechanical loading. This myonuclear accretion is critical for efficient muscle hypertrophy. Herein, we present protocols for the detection of MuSC-derived new myonuclei in loaded mouse muscle, including procedures for EdU injection to stain myonuclei, followed by surgery and skeletal muscle fixation. We then describe immunostaining for EdU+ myonuclei and image acquisition for quantitative analyses. For complete details on the use and execution of this protocol, please refer to Kaneshige et al. (2022). [Display omitted] •Surgical procedure for mechanical loading of the plantaris muscle•Fundamental procedure for preparing frozen muscle samples•Detailed description of immunostaining for EdU+ myonuclei•Acquisition of images for quantitative analyses Publisher's note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. Muscle satellite cells (MuSCs) supply nuclei to existing myofibers in response to mechanical loading. This myonuclear accretion is critical for efficient muscle hypertrophy. Herein, we present protocols for the detection of MuSC-derived new myonuclei in loaded mouse muscle, including procedures for EdU injection to stain myonuclei, followed by surgery and skeletal muscle fixation. We then describe immunostaining for EdU+ myonuclei and image acquisition for quantitative analyses.