Such muscle cell fusion, the researchers say, is not only important for understanding normal muscle growth, but also muscle regeneration after injury or disease. The work, they believe, could further development of therapies for muscular dystrophy or age-related muscle wasting.
Their report on muscle cell cytoskeletons, published in Developmental Cell May 17, adds detail to a previous study last year showing that actin — a main building block of the cell’s cytoskeleton — is required to form those finger-like projections and stimulate muscle cell merges. The new discovery outlines the intricate dance required among cytoskeleton-regulating proteins to precisely construct protrusions that promote muscle cell merging. Specifically, the Johns Hopkins team uncovered the activity of a regulatory protein known as “Blown Fuse,” aptly named because muscle cells lacking this protein fail to fuse.
“Blown Fuse was found to play a role in muscle cell fusion 14 years ago,” says Elizabeth Chen, Ph.D. assistant professor of molecular biology and genetics, “and now we know how Blown Fuse regulates the dynamics of the cytoskeleton to facilitate the invasion of one muscle cell by another.”
The test began with researchers putting fluorescent actin in fruit fly muscle cells that incorporated themselves into the growing actin branches in the finger-like protrusions. Then, the researchers used a laser beam to bleach the fluorescent actin in the region of the finger-like protrusions and waited to see whether and how long it would take for new, unbleached actin to spread from other parts of the cell and be taken up by the growing branches in the “fingers.” In normal muscle cells, it took about two minutes for the fluorescence to return. In muscle cells that lacked Blown Fuse, the fluorescence never fully recovered and the cytoskeleton failed to project finger-like protrusions, probably because the WASP-WIP complex does not come off the ends of the actin filaments to start new actin branches.
“These results suggest that the growing ends of the actin cytoskeleton are occupied by the WASP-WIP protein duo and that without Blown Fuse to dissociate with the WASP-WIP complex and push WASP off the ends, new actin branches cannot be started,” says Chen. “And these shorter and stiffer new branches are critical for generating the finger-like membrane protrusions.”
Through a microscope, the Hopkins team compared the finger-like projections from normal cells with cells lacking Blown Fuse. Normal muscle cells form pointy finger-like protrusions that push into the other muscle cell, but cells without Blown Fuse have fewer and floppier protrusions that don’t push their way in to other muscle cells.
“Modulating the stability of the WASP-WIP complex may represent a general mechanism in regulating cytoskeleton dynamics and generating membrane protrusions,” says Chen.
The study was funded by grants from the National Institutes of Health, the American Heart Association and the Muscular Dystrophy Association.
Other authors of the report are Peng Jin, Rui Duan, Fengbao Luo and Sabrina Hong of the Johns Hopkins University School of Medicine and Guofeng Zhang of the National Institute of Biomedical Imaging and Bioengineering.
Vanessa McMains | EurekAlert!
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