Visualizing sarcomere and cellular dynamics in skeletal muscle to improve cell therapies
Hüttemeister, J.; Rudolph, F.; Radke, M.; Fink, C.; Friedrich, D.; Preibisch, S.; Falcke, M.; Wagner, E.; Lehnart, S. E.; Gotthardt, M.
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The giant striated muscle protein titin integrates into the developing sarcomere to form a stable myofilament system that is extended as myocytes fuse. The logistics underlying myofilament assembly and disassembly have started to emerge with the possibility to follow labeled sarcomere components. Here, we generated the mCherry knock-in at titins Z-disk to study skeletal muscle development and remodeling. We find titins integration into the sarcomere tightly regulated and its unexpected mobility facilitating a homogenous distribution of titin after cell fusion - an integral part of syncytium formation and maturation of skeletal muscle. In adult mCherry-titin mice, treatment of muscle injury by implantation of titin-eGFP myoblasts reveals how myocytes integrate, fuse and contribute to the continuous myofilament system across cell boundaries. Unlike in immature primary cells, titin proteins are retained at the proximal nucleus and do not diffuse across the whole syncytium with implications for future cell-based therapies of skeletal muscle disease.
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