Premium
Visualization of dysferlin‐vesicle dynamics reveals a role for microtubules and kinesin‐motors in dysferlin‐vesicle transport and fusion in skeletal muscle cells
Author(s) -
McDade Joel Ryan,
Michele Daniel
Publication year - 2013
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.27.1_supplement.731.1
Subject(s) - dysferlin , vesicle , vesicle fusion , microbiology and biotechnology , cytoplasm , vesicular transport protein , lipid bilayer fusion , chemistry , microtubule , exocytosis , biology , skeletal muscle , biochemistry , membrane , synaptic vesicle , anatomy
Muscle cells reseal the plasma membrane following wounding, and this process is disrupted in cells lacking dysferlin. Dysferlin‐vesicles accumulate at membrane lesions by an unknown mechanism, and the contribution of dysferlin‐vesicle fusion to resealing is unclear. We used live‐cell imaging of dysferlin‐eGFP expressing L6 myotubes to examine the role of microtubules (MTs) and kinesin motors (KIF5) in dysferlin‐vesicle transport and damage‐induced fusion. Dysferlin‐vesicles colabel with KIF5 and are distributed along microtubules throughout the cytoplasm. Long‐range movement of dysferlin‐vesicles is significantly reduced following pharmacological disruption of MTs, indicating that MTs are required for dysferlin vesicle transport in muscle cells. We demonstrate for the first time that wounding induces fusion of dysferlin‐vesicles to form extremely large cytoplasmic vesicles. Disruption of MTs or expression of dominant‐negative KIF5 inhibited large vesicle formation following wounding, indicating that MT‐based transport of dysferlin‐vesicles is critical for dysferlin‐vesicle fusion. Our data suggest that dysferlin‐vesicles undergo vesicle‐vesicle fusion following wounding, and identify a novel role for MTs and KIF5 in regulating dysferlin‐vesicle function in skeletal muscle cells. This research is supported by the American Heart Association Pre‐doctoral Fellowship