Imaging Approaches to Investigate Myonuclear Positioning in Drosophila
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.
Key wordsDrosophila Muscle Nuclear movement Embryo Larvae
We thank the members of the Baylies Lab for advice, particularly, Krista Dobi, Jonathan Rosen, and Stefanie Windner for providing images for Fig. 3A (Dobi) and 2A Stages 14/15 and Stage 17 (Rosen and Windner, respectively). We also acknowledge our funding agencies, SFRH/BD/52041/2012 (Azevedo), and MDA and NIH NIAMS RO1-068128 (Baylies).
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