Azevedo Mafalda, Schulman Victoria K, Folker Eric, Balakrishnan Mridula, Baylies Mary
Graduate Program in Basic and Applied Biology (GABBA), Institute of Biomedical Sciences Abel Salazar, University of Porto, Porto, Portugal.
Program in Developmental Biology, Sloan Kettering Institute, Baylies Lab, Box 310, 1275 York Avenue, New York, NY, 10065, USA.
Methods Mol Biol. 2016;1411:291-312. doi: 10.1007/978-1-4939-3530-7_19.
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.
在骨骼肌中,细胞核位于每个肌纤维的周边,并沿其长度均匀分布。肌细胞核定位不当与肌肉疾病和肌肉功能下降有关。利用果蝇(黑腹果蝇)已确定了几种调节核位置所需的机制。果蝇和脊椎动物之间肌纤维的保守性、先进遗传工具的可用性以及在体内使用荧光蛋白可视化动态过程的能力,使果蝇成为研究肌细胞核定位的优秀系统。本章描述了使用果蝇胚胎和幼虫进行延时成像和固定成像的方法,以研究肌细胞核定位的机制。