Marine Biological Laboratory, Woods Hole, MA 02543, USA.
J Cell Sci. 2011 Jun 15;124(Pt 12):2086-95. doi: 10.1242/jcs.082263. Epub 2011 May 24.
The mechanical properties of cells change as they proceed through the cell cycle, primarily owing to regulation of actin and myosin II. Most models for cell mechanics focus on actomyosin in the cortex and ignore possible roles in bulk cytoplasm. We explored cell cycle regulation of bulk cytoplasmic actomyosin in Xenopus egg extracts, which is almost undiluted cytoplasm from unfertilized eggs. We observed dramatic gelation-contraction of actomyosin in mitotic (M phase) extract where Cdk1 activity is high, but not in interphase (I-phase) extract. In spread droplets, M-phase extract exhibited regular, periodic pulses of gelation-contraction a few minutes apart that continued for many minutes. Comparing actin nucleation, disassembly and myosin II activity between M-phase and I-phase extracts, we conclude that regulation of nucleation is likely to be the most important for cell cycle regulation. We then imaged F-actin in early zebrafish blastomeres using a GFP-Utrophin probe. Polymerization in bulk cytoplasm around vesicles increased dramatically during mitosis, consistent with enhanced nucleation. We conclude that F-actin polymerization in bulk cytoplasm is cell cycle regulated in early vertebrate embryos and discuss possible biological functions of this regulation.
细胞在细胞周期中经历的机械特性变化主要归因于肌动蛋白和肌球蛋白 II 的调节。大多数细胞力学模型主要集中在皮层中的肌动球蛋白,而忽略了细胞质中可能的作用。我们在爪蟾卵提取物中探索了细胞周期对细胞质肌动球蛋白的调节,该提取物几乎是未受精卵的未稀释细胞质。我们观察到在 Cdk1 活性高的有丝分裂(M 期)提取物中,肌动球蛋白发生剧烈的凝胶收缩,但在间期间期(I 期)提取物中则不会。在展开的液滴中,M 期提取物每隔几分钟就会出现规则的、周期性的凝胶收缩脉冲,持续数分钟。通过比较 M 期和 I 期提取物中的肌动蛋白成核、解聚和肌球蛋白 II 活性,我们得出结论,成核的调节可能是细胞周期调节的最重要因素。然后,我们使用 GFP-Utrophin 探针在早期斑马鱼胚胎细胞中成像 F-肌动蛋白。在有丝分裂期间,囊泡周围的细胞质中 F-肌动蛋白的聚合显著增加,这与成核增强一致。我们得出结论,早期脊椎动物胚胎中细胞质中的 F-肌动蛋白聚合受到细胞周期的调节,并讨论了这种调节的可能生物学功能。