Department of Biology, Duke University, Durham, NC 27708, USA.
Dev Biol. 2010 Sep 15;345(2):117-32. doi: 10.1016/j.ydbio.2010.06.028. Epub 2010 Jun 28.
Metazoan development involves a myriad of dynamic cellular processes that require cytoskeletal function. Nonmuscle myosin II plays essential roles in embryonic development; however, knowledge of its role in post-embryonic development, even in model organisms such as Drosophila melanogaster, is only recently being revealed. In this study, truncation alleles were generated and enable the conditional perturbation, in a graded fashion, of nonmuscle myosin II function. During wing development they demonstrate novel roles for nonmuscle myosin II, including in adhesion between the dorsal and ventral wing epithelial sheets; in the formation of a single actin-based wing hair from the distal vertex of each cell; in forming unbranched wing hairs; and in the correct positioning of veins and crossveins. Many of these phenotypes overlap with those observed when clonal mosaic analysis was performed in the wing using loss of function alleles. Additional requirements for nonmuscle myosin II are in the correct formation of other actin-based cellular protrusions (microchaetae and macrochaetae). We confirm and extend genetic interaction studies to show that nonmuscle myosin II and an unconventional myosin, encoded by crinkled (ck/MyoVIIA), act antagonistically in multiple processes necessary for wing development. Lastly, we demonstrate that truncation alleles can perturb nonmuscle myosin II function via two distinct mechanisms--by titrating light chains away from endogenous heavy chains or by recruiting endogenous heavy chains into intracellular aggregates. By allowing myosin II function to be perturbed in a controlled manner, these novel tools enable the elucidation of post-embryonic roles for nonmuscle myosin II during targeted stages of fly development.
后生动物的发育涉及到许多动态的细胞过程,这些过程都需要细胞骨架的功能。非肌肉肌球蛋白 II 在胚胎发育中起着至关重要的作用;然而,即使在模式生物如黑腹果蝇中,其在胚胎后的发育中的作用的知识,直到最近才被揭示。在这项研究中,产生了截断等位基因,使非肌肉肌球蛋白 II 的功能能够以分级的方式进行条件性扰动。在翅膀发育过程中,它们证明了非肌肉肌球蛋白 II 的新作用,包括在背侧和腹侧翅膀上皮片之间的黏附;在每个细胞的远端顶点从单个基于肌动蛋白的翅膀毛中形成;形成不分叉的翅膀毛;以及正确定位脉和横脉。这些表型中的许多与在翅膀中使用功能丧失等位基因进行克隆嵌合体分析时观察到的表型重叠。非肌肉肌球蛋白 II 的其他要求是在其他基于肌动蛋白的细胞突起(微刚毛和Macrochaetae)的正确形成。我们证实并扩展了遗传相互作用研究,表明非肌肉肌球蛋白 II 和由卷曲(ck/MyoVIIA)编码的非典型肌球蛋白在翅膀发育所必需的多个过程中发挥拮抗作用。最后,我们证明截断等位基因可以通过两种不同的机制来干扰非肌肉肌球蛋白 II 的功能——通过从内源性重链上滴定轻链或通过将内源性重链募集到细胞内聚集体中。通过允许以受控的方式干扰肌球蛋白 II 的功能,这些新工具使我们能够在苍蝇发育的靶向阶段阐明非肌肉肌球蛋白 II 的胚胎后作用。