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Immunolabelling Myofiber Degeneration in Muscle Biopsies
Published on: December 5, 2019
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Improved immunoblotting methods provide critical insights into phenotypic differences between two murine
Amber L Mueller1, Patrick F Desmond, Ru-Ching Hsia
1University of Maryland School of Medicine, Department of Physiology, Baltimore, Maryland, 21201, USA.
Muscle & Nerve
|March 19, 2014
Summary
Proteomics revealed endoplasmic reticulum (ER) stress is elevated in A/J mice, suggesting it underlies phenotypic differences in murine dysferlinopathy models and potentially human diseases.
Area of Science:
- Muscle Biology
- Proteomics
- Disease Modeling
Background:
- Dysferlinopathies are genetic muscle-wasting diseases.
- A/J and B10.SJL mice are established models for studying dysferlinopathies.
- Phenotypic differences exist between these murine models.
Purpose of the Study:
- To investigate phenotypic differences between A/J and B10.SJL murine dysferlinopathy models using proteomics.
- To identify molecular mechanisms contributing to disease variability.
Main Methods:
- Optimized immunoblotting techniques for dysferlin detection in tibialis anterior (TA) muscle homogenates.
- Compared protein levels of dysferlin, its interacting partners (MG53, annexin-A2, caveolin-3), and ER stress marker CHOP in control, A/J, and B10.SJL mice.
- Utilized immunoelectron microscopy to determine the subcellular localization of dysferlin.
Main Results:
- Radioimmunoprecipitation assay (RIPA) buffer and sonication enhanced dysferlin immunoblotting.
- Elevated levels of the ER stress marker CHOP were observed in A/J mouse muscle.
- Dysferlin was localized to membranes near the Z-disk, associated with Golgi, ER, and sarcoplasmic reticulum (SR) networks.
Conclusions:
- Endoplasmic reticulum (ER) stress may explain phenotypic variations between A/J and B10.SJL murine dysferlinopathy models.
- ER stress could be a contributing factor in human dysferlinopathies.
