Role of the centrosome in organizing the interphase microtubule array: properties of cytoplasts containing or lacking centrosomes.
Role of the centrosome in organizing the interphase microtubule array: properties of cytoplasts containing or lacking centrosomes.
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DOI：
10.1083/jcb.98.5.1763
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发表时间：
1984-05
影响因子：
7.8
通讯作者：
Kirschner, M
中科院分区：
文献类型：
--
作者：
Karsenti, E;Kobayashi, S;Mitchison, T;Kirschner, M
为了研究中心体在间期细胞微管组织中的作用，我们开发了一种获得含有或不含有中心体的细胞质(无核细胞)的方法。在药物诱导的微管解聚后，由稀疏的板状细胞制成的带有中心体的细胞质重新构成了典型的正常细胞的微管阵列。在此条件下，没有中心体的细胞质仅形成少量分散的微管。这种聚合度的差异表明，中心体不仅影响细胞中微管的分布，而且影响微管的数量。令我们惊讶的是，微管的组装程度随着原始培养物中细胞密度的增加而增加。在融合密度下，没有中心体的细胞质有许多微管，相当于有中心体的细胞质。这些额外的微管排列在外围，与中心体微管不同，它们对诺可唑的敏感性不同。这些和其他结果表明，中心体稳定了细胞中的微管，可能是通过覆盖一端。对诺可达唑更敏感的微管是由于细胞生长状态的改变而产生的，可能代表自由或无封端的聚合物。这些实验表明，微管的空间排列可能会通过改变微管的总浓度或组装的临界浓度来改变。
To study the role of the centrosome in microtubule organization in interphase cells, we developed a method for obtaining cytoplasts (cells lacking a nucleus) that did or did not contain centrosomes. After drug- induced microtubule depolymerization, cytoplasts with centrosomes made from sparsely plated cells reconstituted a microtubule array typical of normal cells. Under these conditions cytoplasts without centrosomes formed only a few scattered microtubules. This difference in degree of polymerization suggests that centrosomes affect not only the distribution but the amount of microtubules in cells. To our surprise, the extent of microtubules assembled increased with the cell density of the original culture. At confluent density, cytoplasts without centrosomes had many microtubules, equivalent to cytoplasts with centrosomes. The additional microtubules were arranged peripherally and differed from the centrosomal microtubules in their sensitivity to nocodazole. These and other results suggest that the centrosome stabilizes microtubules in the cell, perhaps by capping one end. Microtubules with greater sensitivity to nocodazole arise by virtue of change in the growth state of the cell and may represent free or uncapped polymers. These experiments suggest that the spatial arrangement of microtubules may change by shifting the total tubulin concentration or the critical concentration for assembly.