Improved immunoblotting methods provide critical insights into phenotypic differences between two murine dysferlinopathy models.

Abstract:

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.

journal_name

Muscle Nerve

journal_title

Muscle & nerve

authors

Mueller AL,Desmond PF,Hsia RC,Roche JA

doi

10.1002/mus.24220

subject

Has Abstract

pub_date

2014-08-01 00:00:00

pages

286-9

issue

2

eissn

0148-639X

issn

1097-4598

journal_volume

50

pub_type

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