REAL-TIME VISUALIZATION OF CELL-CYCLE DEPENDENT CHANGES IN MICROTUBULE DYNAMICS IN CYTOPLASMIC EXTRACTS
REAL-TIME VISUALIZATION OF CELL-CYCLE DEPENDENT CHANGES IN MICROTUBULE DYNAMICS IN CYTOPLASMIC EXTRACTS
复制标题
DOI：
10.1016/0092-8674(90)90022-7
复制
发表时间：
1990-08-10
期刊：
影响因子：
64.5
通讯作者：
MITCHISON, TJ
中科院分区：
文献类型：
--
作者：
BELMONT, LD;HYMAN, AA;MITCHISON, TJ
使用爪蟾卵提取物逮捕间期或有丝分裂，我们直接观察到微管动力学在细胞周期的不同阶段的差异。间期提取物从减数分裂后第一个间期的卵中制备。通过将纯化的细胞周期蛋白添加到间期提取物中来制备有丝分裂提取物。微管成核的中心体和可视化的荧光显微镜提取物中加入罗丹明标记的微管蛋白。 我们发现有丝分裂与间期提取物中微管动力学的显着差异。定量分析表明，聚合和解聚的速率是相似的，在间期和有丝分裂，在我们的实验的空间和时间分辨率的动力学的差异几乎完全是由于在有丝分裂提取物的增长到收缩（灾难频率）的过渡频率的增加。
Using Xenopus egg extracts arrested in interphase or mitosis, we directly observed differences in microtubule dynamics at different stages of the cell cycle. Interaphase extracts were prepared from eggs in the first interphase after meiosis. Mitotic extracts were prepared by addition of purified cyclin to interphase extracts. Microtubules were nucleated by the addition of centrosomes and visualized by fluorescence videomicroscopy in extracts to which rhodamine-labeled tubulin had been added. We found a striking difference in microtubule dynamics in mitotic versus interphase extracts. Quantitative analysis revealed that the rates of polymerization and depolymerization are similar in interphase and mitosis and that within the spatial and temporal resolution of our experiments the difference in dynamics is due almost entirely to an increase in the frequency of transition from growing to shrinking (catastrophe frequency) in the mitotic extracts.