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Updated: Jan 1, 2026
Immunolabelling Myofiber Degeneration in Muscle Biopsies
Published on: December 5, 2019
Immunolabelling Myofiber Degeneration in Muscle Biopsies
Maximilien Bencze1, Baptiste Periou2, Yasmine Baba-Amer2
1Inserm, Institut Mondor de Recherche Biomédicale; The Dubowitz Neuromuscular Centre, Molecular Neurosciences Section, Developmental Neurosciences Program, UCL Great Ormond Street Institute of Child Health; m.bencze@ucl.ac.uk.
Abstract:
The necrosis of muscle fibres (myonecrosis) plays a central role in the pathogenesis of several muscle conditions, including muscular dystrophies. Therapeutic options addressing the causes of muscular dystrophy pathogenesis are expected to alleviate muscle degeneration. Therefore, a method to assay and quantify the extent of cell death in muscle biopsies is needed. Conventional methods to observe myofiber degeneration in situ are either poorly quantitative or rely on the injection of vital dyes. In this article, an immunofluorescence protocol is described that stains necrotic myofibers by targeting immunoglobulin G (IgG) uptake by myofibers. The IgG uptake method is based on cell features characterizing the necrotic demise, including 1) the loss of plasma membrane integrity with the release of damage-associated molecular patterns and 2) the uptake of plasmatic proteins. In murine cross-sections, the co-immunolabelling of myofibers, extracellular matrix proteins, and mouse IgG allows clean and straightforward identification of myofibers with necrotic fate. This simple method is suitable for quantitative analysis and applicable to all species, including human samples, and does not require the injection of vital dye. The staining of necrotic myofibers by IgG uptake can also be paired with other co-immunolabelling.
Insights
A new immunofluorescence protocol quantifies muscle fiber necrosis by tracking immunoglobulin G (IgG) uptake. This method accurately identifies myonecrosis in muscle biopsies without vital dyes, aiding muscular dystrophy research.
Area of Science:
- Muscle biology
- Pathology
- Immunohistochemistry
Background:
- Myonecrosis is central to muscular dystrophies and other muscle conditions.
- Assessing muscle fiber death is crucial for developing effective therapies.
- Current methods for quantifying myonecrosis are often imprecise or invasive.
Purpose of the Study:
- To develop a quantitative and reliable method for assaying myonecrosis in muscle biopsies.
- To establish an immunofluorescence protocol for identifying necrotic muscle fibers.
Main Methods:
- Described an immunofluorescence protocol targeting immunoglobulin G (IgG) uptake by necrotic muscle fibers.
- Utilized co-immunolabeling of myofibers, extracellular matrix proteins, and mouse IgG in murine cross-sections.
- Protocol relies on loss of plasma membrane integrity and plasmatic protein uptake characteristic of necrotic cells.
Main Results:
- The IgG uptake method allows for clean and straightforward identification of necrotic myofibers.
- Demonstrated suitability for quantitative analysis in murine samples.
- Method is applicable across species, including human samples, without vital dye injection.
Conclusions:
- Developed a simple, quantitative immunofluorescence protocol for identifying necrotic muscle fibers via IgG uptake.
- This method provides a valuable tool for studying myonecrosis in various muscle conditions.
- The protocol is versatile and can be combined with other co-immunolabeling techniques.
