Royou Anne, Sullivan William, Karess Roger
Centre National de la Recherche Scientifique, Centre de Génétique Moléculaire, 91198 Gif-sur-Yvette, France.
J Cell Biol. 2002 Jul 8;158(1):127-37. doi: 10.1083/jcb.200203148.
The nuclei of early syncytial Drosophila embryos migrate dramatically toward the poles. The cellular mechanisms driving this process, called axial expansion, are unclear, but myosin II activity is required. By following regulatory myosin light chain (RLC)-green fluorescent protein dynamics in living embryos, we observed cycles of myosin recruitment to the cortex synchronized with mitotic cycles. Cortical myosin is first seen in a patch at the anterocentral part of the embryo at cycle 4. With each succeeding cycle, the patch expands poleward, dispersing at the beginning of each mitosis and reassembling at the end of telophase. Each cycle of actin and myosin recruitment is accompanied by a cortical contraction. The cortical myosin cycle does not require microtubules but correlates inversely with Cdc2/cyclinB (mitosis-promoting factor) activity. A mutant RLC lacking inhibitory phosphorylation sites was fully functional with no effect on the cortical myosin cycle, indicating that Cdc2 must be modulating myosin activity by some other mechanism. An inhibitor of Rho kinase blocks the cortical myosin recruitment cycles and provokes a concomitant failure of axial expansion. These studies suggest a model in which cycles of myosin-mediated contraction and relaxation, tightly linked to Cdc2 and Rho kinase activity, are directly responsible for the axial expansion of the syncytial nuclei.
早期合胞体果蝇胚胎的细胞核会剧烈地向两极迁移。驱动这一过程(称为轴向扩张)的细胞机制尚不清楚,但肌球蛋白II的活性是必需的。通过追踪活胚胎中调节性肌球蛋白轻链(RLC)-绿色荧光蛋白的动态变化,我们观察到肌球蛋白向皮质募集的周期与有丝分裂周期同步。皮质肌球蛋白在第4周期时首次出现在胚胎前中央部分的一个斑块中。随着每个后续周期,该斑块向两极扩展,在每次有丝分裂开始时分散,并在末期结束时重新组装。肌动蛋白和肌球蛋白募集的每个周期都伴随着一次皮质收缩。皮质肌球蛋白周期不需要微管,但与Cdc2/细胞周期蛋白B(有丝分裂促进因子)的活性呈负相关。缺乏抑制性磷酸化位点的突变型RLC功能完全正常,对皮质肌球蛋白周期没有影响,这表明Cdc2必须通过其他机制调节肌球蛋白的活性。Rho激酶的抑制剂会阻断皮质肌球蛋白的募集周期,并引发轴向扩张的同时失败。这些研究提出了一个模型,其中肌球蛋白介导的收缩和舒张周期与Cdc2和Rho激酶的活性紧密相连,直接导致合胞体细胞核的轴向扩张。