This work was funded by a grant to J.A.R. from the Jain Foundation Inc.
Improved immunoblotting methods provide critical insights into phenotypic differences between two murine dysferlinopathy models
Version of Record online: 14 JUL 2014
Copyright © 2014 Wiley Periodicals, Inc.
Muscle & Nerve
Volume 50, Issue 2, pages 286–289, August 2014
How to Cite
Mueller, A. L., Desmond, P. F., Hsia, R.-c. and Roche, J. A. (2014), Improved immunoblotting methods provide critical insights into phenotypic differences between two murine dysferlinopathy models. Muscle Nerve, 50: 286–289. doi: 10.1002/mus.24220
The authors declare no conflicts of interest.
- Issue online: 21 JUL 2014
- Version of Record online: 14 JUL 2014
- Accepted manuscript online: 23 FEB 2014 04:15AM EST
- Manuscript Accepted: 20 FEB 2014
- dysferlin binding proteins;
- ER stress;
- skeletal muscle
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. Muscle Nerve 50:286–289, 2014