SELECTIVE STABILIZATION OF MICROTUBULES ORIENTED TOWARD THE DIRECTION OF CELL-MIGRATION
SELECTIVE STABILIZATION OF MICROTUBULES ORIENTED TOWARD THE DIRECTION OF CELL-MIGRATION
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DOI：
10.1073/pnas.85.16.5946
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发表时间：
1988-08-01
影响因子：
11.1
通讯作者：
BULINSKI, JC
中科院分区：
文献类型：
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作者：
GUNDERSEN, GG;BULINSKI, JC
在许多培养的细胞中，微管（MT）阵列的一小部分不表现出快速的周转（t1/2 approxeq. 10分钟）显示的大多数细胞MT。MT的稳定类的功能是未知的，并且已经被细胞内稳定MT的明显缺乏组织所混淆。使用针对去酪氨酸微管蛋白的抗体，微管蛋白的后修饰形式，积累在稳定的MT，我们本地化的稳定MT在小鼠3 T3细胞诱导开始定向迁移的实验创伤的汇合单层。单层创伤后，立即在伤口边缘的细胞中的稳定的MT的分布类似于在单层内部的细胞中，大多数细胞中含有随机分布的稳定的MT或完全缺乏它们。然而，在创伤后20分钟，在伤口边缘的细胞开始产生不对称的MT阵列，几乎所有稳定的MT都朝向与伤口接触的细胞边缘。单层损伤后2小时，伤口边缘80%的细胞产生了这种稳定MT的极化阵列，并且该阵列保持至少12小时。极化阵列中的MT显示出对诺考达唑解聚的增强抗性，从而提供了对其稳定性的独立测试。形成的极性阵列稳定的MT出现之前发病的细胞迁移和密切相关的MT组织中心的重新定位。这些结果表明，培养的细胞可以重塑他们的MT阵列迅速响应于细胞外信号，并建议，选择性稳定的MT是一个早期事件中产生的细胞不对称性。
A small subset of the microtubule (MT) array in many cultured cells does not exhibit the rapid turnover (t1/2 .apprxeq. 10 min) shown by most cellular MTs. The function of the stable class of MTs is unknown and has been confounded by the apparent lack of organization of stable MTs within cells. Using an antibody against detyrosinated tubulin, a post-translationally modified form of tubulin that accumulates in stable MTs, we localized the stable MTs in mouse 3T3 cells induced to initiate directional migration by experimental wounding of confluent monolayers. Immediately after monolayer wounding, the distribution of stable MTs in cells at the wound edge resembled that in cells in the monolayer interior; most cells either contained randomly distributed stable MTs or lacked them entirely. However, by 20 min after wounding, cells at the wound margin began to generate an asymmetric MT array, with virtually all stable MTs oriented toward the cell edge in contact with the wound. Two hours after monolayer wounding, .gtoreq. 80% of cells at the wound margin had generated this polarized array of stable MTs, and the array was maintained for at least 12 hr. MTs in the polarized array showed enhanced resistance to depolymerization by nocodazole, thus providing an independent test of their stability. Formation of the polar array of stable MTs appeared to precede onset of cell migration and closely paralleled reorientation of the MT-organizing center. These results show that cultured cells can remodel their MT array rapidly in response to an extracellular signal and suggest that selective stabilization of MTs is an early event in the generation of cellular asymmetry.