Whole-mount immunofluorescence staining of mesenchymal progenitors in murine plantaris muscle

Tamaki Kurosawa; Madoka Ikemoto-Uezumi; Akihiro Kaneshige; So-Ichiro Fukada; Akiyoshi Uezumi

doi:10.1016/j.xpro.2022.101593

Whole-mount immunofluorescence staining of mesenchymal progenitors in murine plantaris muscle

STAR Protoc. 2022 Jul 31;3(3):101593. doi: 10.1016/j.xpro.2022.101593. eCollection 2022 Sep 16.

Authors

Tamaki Kurosawa¹, Madoka Ikemoto-Uezumi², Akihiro Kaneshige³, So-Ichiro Fukada⁴, Akiyoshi Uezumi⁵

Affiliations

¹ Department of Nutritional Physiology, Institute of Medical Nutrition, Tokushima University Graduate School, 3-18-15 Kuramoto-cho, Tokushima 770-8503, Japan; Laboratory of Veterinary Pharmacology, Department of Veterinary Medical Sciences, Graduate School of Agriculture and Life Sciences, Tokyo University, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan.
² Department of Nutritional Physiology, Institute of Medical Nutrition, Tokushima University Graduate School, 3-18-15 Kuramoto-cho, Tokushima 770-8503, Japan.
³ Project for Muscle Stem Cell Biology, Graduate School of Pharmaceutical Sciences, Osaka University, 1-6 Yamada-oka, Suita, Osaka 565-0871, Japan; Biological/Pharmacological Research Laboratories, Central Pharmaceutical Research Institute, Japan Tobacco Inc., 1-1 Murasaki-cho, Takatsuki, Osaka 569-1125, Japan; Laboratory of Molecular and Cellular Physiology, Graduate School of Pharmaceutical Sciences, Osaka University, 1-6 Yamada-oka, Suita, Osaka 565-0871, Japan.
⁴ Project for Muscle Stem Cell Biology, Graduate School of Pharmaceutical Sciences, Osaka University, 1-6 Yamada-oka, Suita, Osaka 565-0871, Japan.
⁵ Department of Nutritional Physiology, Institute of Medical Nutrition, Tokushima University Graduate School, 3-18-15 Kuramoto-cho, Tokushima 770-8503, Japan. Electronic address: uezumi.akiyoshi@tokushima-u.ac.jp.

Abstract

We recently demonstrated that mesenchymal progenitors play a critical role in regulating satellite cell-dependent myonuclear accretion during overload-induced muscle hypertrophy. Here, we describe the detailed protocol for whole-mount immunofluorescence staining of mesenchymal progenitors in mouse plantaris muscle. Z-stack image reconstruction provides a whole-cell image and enables examination of YAP nuclear translocation in mesenchymal progenitors induced by overload. For complete details on the use and execution of this protocol, please refer to Kaneshige et al. (2022a).

Keywords: Antibody; Microscopy; Model Organisms; Stem Cells.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Fluorescent Antibody Technique
Mice
Muscle, Skeletal / physiology
Satellite Cells, Skeletal Muscle* / physiology
Staining and Labeling