Snyder J A, McLelland S L
Department of Biological Sciences, University of Denver, CO 80208, USA.
Cell Biol Int. 1996 Aug;20(8):573-8. doi: 10.1006/cbir.1996.0075.
Mitotic PtK1 cells were treated both during mid-anaphase and at furrow initiation with the potent microtubule (MT) stabilizing agent, taxol, to determine the role of MTs in the rate of cytokinetic events. Rates of cytokinesis (micron/min) were measured by changes in furrow diameter. Incubation of PtK1 cells during mid-anaphase with 5 micrograms/ml taxol slows the rate of cytokinesis by an average of 43%. Instead of furrow initiation to midbody formation taking an average of 10.7 min (1.6 microns/min), furrowing to midbody formation was completed in an average of 19.0 min (0.9 micron/min), which does not include the 7-min period between taxol application in mid-anaphase and furrow initiation. Application of 5 micrograms/ml taxol to cells at furrow initiation had a reduced effect on decreasing the rate of cytokinesis and midbody formation; furrowing to midbody formation took an average of 14.6 min (1.2 microns/min). These data suggest that delays in the rate of cytokinesis is dependent on the mitotic stage at which taxol is applied. Ultrastructural analysis shows that taxol treatment of anaphase cells prevents midbody formation during early G1, yet MT number and organization in the furrowed region is not significantly altered from untreated cells. There is little change in the organization and amount of contractile ring microfilaments, yet filaments are also found parallel to midbody MTs. Our results may be explained by the fact that taxol tends to stabilize MTs which probably affects the rate at which they depolymerize in the terminal phases of cytokinesis. Reduction in depolymerization rates of a stable population of MTs could serve to regulate the rate of cytokinesis.
用强效微管(MT)稳定剂紫杉醇处理有丝分裂的PtK1细胞,处理时间为中期后期和沟起始期,以确定微管在胞质分裂事件速率中的作用。通过沟直径的变化来测量胞质分裂速率(微米/分钟)。在中期后期用5微克/毫升紫杉醇孵育PtK1细胞,胞质分裂速率平均减慢43%。从沟起始到中间体形成平均需要10.7分钟(1.6微米/分钟),而现在从沟形成到中间体形成平均需要19.0分钟(0.9微米/分钟),这还不包括在中期后期施加紫杉醇到沟起始之间的7分钟时间。在沟起始时向细胞施加5微克/毫升紫杉醇,对降低胞质分裂速率和中间体形成的影响较小;从沟形成到中间体形成平均需要14.6分钟(1.2微米/分钟)。这些数据表明,胞质分裂速率的延迟取决于紫杉醇施加的有丝分裂阶段。超微结构分析表明,用紫杉醇处理后期细胞可防止在G1早期形成中间体,但沟区域的微管数量和组织与未处理细胞相比没有显著改变。收缩环微丝的组织和数量变化不大,但也发现微丝与中间体微管平行。我们的结果可能可以这样解释:紫杉醇倾向于稳定微管,这可能会影响它们在胞质分裂末期解聚的速率。稳定微管群体解聚速率的降低可能有助于调节胞质分裂的速率。