SUMMARY
Sarcomeres, the fundamental contractile units of muscles, are conserved structures composed of actin thin filaments and myosin thick filaments. How sarcomeres are formed and maintained is not well understood. Here, we show that knockdown of
Drosophila
cofilin (
DmCFL
), an actin depolymerizing factor, disrupts both sarcomere structure and muscle function. The loss of
DmCFL
also results in the formation of sarcomeric protein aggregates and impairs sarcomere addition during growth. The activation of the proteasome delays muscle deterioration in our model. Furthermore, we investigate how a point mutation in CFL2 that causes nemaline myopathy (NM) in humans affects CFL function and leads to the muscle phenotypes observed
in vivo
. Our data provide significant insights to the role of CFLs during sarcomere formation, as well as mechanistic implications for disease progression in NM patients.
In the skeletal muscle, nuclei are positioned at the periphery of each myofiber and are evenly distributed along its length. Improper positioning of myonuclei has been correlated with muscle disease and decreased muscle function. Several mechanisms required for regulating nuclear position have been identified using the fruit fly, Drosophila melanogaster. The conservation of the myofiber between the fly and vertebrates, the availability of advanced genetic tools, and the ability to visualize dynamic processes using fluorescent proteins in vivo makes the fly an excellent system to study myonuclear positioning. This chapter describes time-lapse and fixed imaging methodologies using both the Drosophila embryo and the larva to investigate mechanisms of myonuclear positioning.
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