The role of cytoskeleton in organizing growth cones: a microfilament-associated growth cone component depends upon microtubules for its localization.
The role of cytoskeleton in organizing growth cones: a microfilament-associated growth cone component depends upon microtubules for its localization.
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DOI：
10.1083/jcb.109.4.1621
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发表时间：
1989-10
期刊：
影响因子：
--
通讯作者：
Solomon F
中科院分区：
文献类型：
--
作者：
Goslin K;Birgbauer E;Banker G;Solomon F
我们感兴趣的是细胞骨架和极化细胞形态的组织之间的关系。我们在这里显示，海马神经元的生长锥在文化中被称为13 H9的单克隆抗体特异性染色。在其他系统中，13 H9识别的抗原与鸡红细胞的边缘带相关，并显示微管和微丝相关蛋白的性质（Birgbauer，E.，和F.所罗门。1989 J. Cell Biol.109：1609-1620）。这种双重性质在海马神经元中也很明显。在接种后的早期阶段，抗体染色介导初始细胞铺展的周向板状伪足。随着过程的出现，13 H9染色大量集中在生长锥的远端区域，特别是在板状伪足扇。在这些细胞中，13 H9染色与组装的微管的定位互补。它与组装的肌动蛋白的鬼笔环肽染色部分共定位，但不完全共定位。与诺考达唑孵育快速诱导微管解聚，其在该过程中以远端至近端方向进行。与此同时，13 H9染色发生了快速而显著的重新分布;它沿沿着轴突轴离域，变得与鬼笔环肽染色明显不同，并且始终保持在组装微管后退前端的远端。在没有组装的微管的较长时间后，不能检测到13 H9的染色。去除诺考达唑允许微管以有序的近端到远端的方式改革。13 H9免疫反应性也再次出现，但仅在生长锥中，而不是在沿轴突的任何中间位置沿着，并且仅在微管的重新形成完成之后。结果表明，13 H9识别的抗原是高度集中在生长锥，与聚合肌动蛋白密切相关，其适当的定位取决于完整的微管。
We are interested in the relationship between the cytoskeleton and the organization of polarized cell morphology. We show here that the growth cones of hippocampal neurons in culture are specifically stained by a monoclonal antibody called 13H9. In other systems, the antigen recognized by 13H9 is associated with marginal bands of chicken erythrocytes and shows properties of both microtubule-and microfilament- associated proteins (Birgbauer, E., and F. Solomon. 1989 J. Cell Biol. 109:1609-1620). This dual nature is manifest in hippocampal neurons as well. At early stages after plating, the antibody stains the circumferential lamellipodia that mediate initial cell spreading. As processes emerge, 13H9 staining is heavily concentrated in the distal regions of growth cones, particularly in lamellipodial fans. In these cells, the 13H9 staining is complementary to the localization of assembled microtubules. It colocalizes partially, but not entirely, with phalloidin staining of assembled actin. Incubation with nocodazole rapidly induces microtubule depolymerization, which proceeds in the distal-to-proximal direction in the processes. At the same time, a rapid and dramatic redistribution of the 13H9 staining occurs; it delocalizes along the axon shaft, becoming clearly distinct from the phalloidin staining and always remaining distal to the receding front of assembled microtubules. After longer times without assembled microtubules, no staining of 13H9 can be detected. Removal of the nocodazole allows the microtubules to reform, in an ordered proximal-to- distal fashion. The 13H9 immunoreactivity also reappears, but only in the growth cones, not in any intermediate positions along the axon, and only after the reformation of microtubules is complete. The results indicate that the antigen recognized by 13H9 is highly concentrated in growth cones, closely associated with polymerized actin, and that its proper localization depends upon intact microtubules.