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Sarcolemmal Repair Is a Slow Process and Includes EHD2
Author(s) -
Marg Andreas,
Schoewel Verena,
Timmel Tobias,
Schulze Anne,
Shah Claudio,
Daumke Oliver,
Spuler Simone
Publication year - 2012
Publication title -
traffic
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.677
H-Index - 130
eISSN - 1600-0854
pISSN - 1398-9219
DOI - 10.1111/j.1600-0854.2012.01386.x
Subject(s) - microbiology and biotechnology , biology , myogenesis , dysferlin , annexin , actin , membrane protein , skeletal muscle , cell , myocyte , membrane , anatomy , biochemistry
Skeletal muscle is continually subjected to microinjuries that must be repaired to maintain structure and function. Fluorescent dye influx after laser injury of muscle fibers is a commonly used assay to study membrane repair. This approach reveals that initial resealing only takes a few seconds. However, by this method the process of membrane repair can only be studied in part and is therefore poorly understood. We investigated membrane repair by visualizing endogenous and GFP ‐tagged repair proteins after laser wounding. We demonstrate that membrane repair and remodeling after injury is not a quick event but requires more than 20 min. The endogenous repair protein dysferlin becomes visible at the injury site after 20 seconds but accumulates further for at least 30 min. Annexin A1 and F ‐actin are also enriched at the wounding area. We identified a new participant in the membrane repair process, the ATPase EHD2 . We show, that EHD2 , but not EHD1 or mutant EHD2 , accumulates at the site of injury in human myotubes and at a peculiar structure that develops during membrane remodeling, the repair dome. In conclusion, we established an approach to visualize membrane repair that allows a new understanding of the spatial and temporal events involved.