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Updated: May 2, 2026
Immunolabelling Myofiber Degeneration in Muscle Biopsies
Published on: December 5, 2019
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.
Introduction:
We adopted a proteomics-based approach to gain insights into phenotypic differences between A/J and B10.SJL murine dysferlinopathy models.
Methods:
We optimized immunoblotting of dysferlin by preparing homogenates of the tibialis anterior (TA) muscle under several different conditions. We compared TA muscles of control, A/J, and B10.SJL mice for levels of dysferlin; dysferlin's partners MG53, annexin-A2, and caveolin-3; and the endoplasmic reticulum (ER) stress marker CHOP. We performed immunoelectron microscopy on control rat TA muscle to determine the precise location of dysferlin.
Results:
RIPA (radioimmunoprecipitation assay) buffer and sonication improves immunoblotting of dysferlin. The ER stress marker CHOP is elevated in A/J muscle. Dysferlin is localized mostly to membranes close to the Z-disk that have been reported to be part of the Golgi, ER, and sarcoplasmic reticulum (SR) networks.
Conclusions:
ER stress might underlie phenotypic differences between A/J and B10.SJL mice and play a role in human dysferlinopathies.
