Sellitto C, Kuriyama R
Department of Cell Biology and Neuroanatomy, University of Minnesota, Minneapolis 55455.
J Cell Sci. 1988 Jan;89 ( Pt 1):57-65. doi: 10.1242/jcs.89.1.57.
Mitotic Chinese hamster ovary cells were obtained by treatment with microtubule drugs under various conditions, and the shape of spindles was analysed by phase-contrast microscopy of isolated spindles, and by indirect immunofluorescence staining of whole mitotic cells with anti-tubulin antibody. Bipolarity of spindles was maintained after treatment with 0.05 microM of colcemid for 3.5 h, but increased exposure to higher concentrations (0.32 microM) and for longer durations (5.5 h) led to a marked rise in multipolar spindles. Nocodazole treatment, on the other hand, failed to show a multiplicity of spindle poles even at 3.3 microM. Each pole of a multipolar spindle was associated with pericentriolar material, as shown by staining with an autoimmune serum specific for pericentriolar material. The number of locations with free pericentriolar material capable of polymerizing microtubules in vitro also increased with increasing numbers of spindle poles, suggesting that dispersion of the pericentriolar material resulted in the production of many microtubule-nucleating sites in multipolar spindles. The different efficiencies of recovery from different drugs, which have been known to be quite variable, may be partly due to the different extent of dispersion of the pericentriolar material.
通过在不同条件下用微管药物处理获得有丝分裂的中国仓鼠卵巢细胞，通过对分离的纺锤体进行相差显微镜观察，以及用抗微管蛋白抗体对整个有丝分裂细胞进行间接免疫荧光染色来分析纺锤体的形状。用0.05微摩尔秋水仙酰胺处理3.5小时后纺锤体的双极性得以维持，但暴露于更高浓度（0.32微摩尔）更长时间（5.5小时）会导致多极纺锤体显著增加。另一方面，即使在3.3微摩尔的浓度下，诺考达唑处理也未显示出纺锤体极的多样性。如用针对中心粒外周物质的自身免疫血清染色所示，多极纺锤体的每个极都与中心粒外周物质相关。体外能够聚合微管的游离中心粒外周物质的位置数量也随着纺锤体极数量的增加而增加，这表明中心粒外周物质的分散导致多极纺锤体中产生许多微管成核位点。已知不同药物恢复的效率差异很大，这可能部分归因于中心粒外周物质分散程度的不同。
