Cytoplasmic dynein and LIS1 are required for microtubule advance during growth cone remodeling and fast axonal outgrowth
Cytoplasmic dynein and LIS1 are required for microtubule advance during growth cone remodeling and fast axonal outgrowth
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DOI：
10.1523/jneurosci.1135-07.2007
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发表时间：
2007-05-23
影响因子：
5.3
通讯作者：
Vallee, Richard B.
中科院分区：
文献类型：
--
作者：
Grabham, Peter W.;Seale, Garrett E.;Vallee, Richard B.
最近的证据表明动力蛋白及其调节因子 dynactin 和 LIS1 与神经元和非神经元细胞迁移有关。在当前的研究中，我们试图测试对神经元细胞运动的影响是否可能部分反映这些蛋白质在生长锥中的作用。在接受急性层粘连蛋白处理的小鸡感觉神经元中，动力蛋白、动力蛋白和 LIS1 被惊人且快速地动员到生长锥的前缘，在那里它们被发现与会聚成层粘连蛋白诱导的轴突生长的微管相关。为了强烈干扰 LIS1 和动力蛋白功能并尽量减少继发表型效应，我们在轴突启动前注射了针对这些蛋白质的抗体。这两种蛋白的抗体几乎完全阻断了层粘连蛋白诱导的生长锥重塑和潜在的微管重组。根据增强型绿色荧光蛋白微管蛋白和微管尖端相关 EB3（末端结合蛋白 3）的实时分析判断，注射抗体显着降低了微管对分化轴突生长锥周围区域的渗透。动力蛋白和 LIS1 抑制对微管组装没有可检测到的影响，但降低了微管抵抗逆行肌动蛋白流的能力。在海马神经元中，动力蛋白、动力蛋白和 LIS1 在第 3 阶段在轴突生长锥中富集，并且生长锥组织和轴突伸长都因 LIS1 RNA 干扰而改变。总之，我们的数据表明，动力蛋白和 LIS1 在与轴突发生相关的生长锥重塑过程中的微管前进中发挥着令人惊讶的突出作用。这些数据可以部分解释这些蛋白质在大脑发育疾病中的作用，并支持在神经元分化和神经系统发育的各个方面的重要作用。
Recent evidence has implicated dynein and its regulatory factors dynactin and LIS1 in neuronal and non-neuronal cell migration. In the current study we sought to test whether effects on neuronal cell motility might reflect, in part, a role for these proteins in the growth cone. In chick sensory neurons subjected to acute laminin treatment dynein, dynactin, and LIS1 were mobilized strikingly and rapidly to the leading edge of the growth cone, where they were seen to be associated with microtubules converging into the laminin-induced axonal outgrowths. To interfere acutely with LIS1 and dynein function and to minimize secondary phenotypic effects, we injected antibodies to these proteins just before axon initiation. Antibody to both proteins produced an almost complete block of laminin-induced growth cone remodeling and the underlying reorganization of microtubules. Penetration of microtubules into the peripheral zone of differentiating axonal growth cones was decreased dramatically by antibody injection, as judged by live analysis of enhanced green fluorescent proteintubulin and the microtubule tip-associated EB3 (end-binding protein 3). Dynein and LIS1 inhibition had no detectable effect on microtubule assembly but reduced the ability of microtubules to resist retrograde actin flow. In hippocampal neurons dynein, dynactin, and LIS1 were enriched in axonal growth cones at stage 3, and both growth cone organization and axon elongation were altered by LIS1 RNA interference. Together, our data indicate that dynein and LIS1 play a surprisingly prominent role in microtubule advance during growth cone remodeling associated with axonogenesis. These data may explain, in part, the role of these proteins in brain developmental disease and support an important role in diverse aspects of neuronal differentiation and nervous system development.