Neuronal polarity selection by topography-induced focal adhesion control
Neuronal polarity selection by topography-induced focal adhesion control
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DOI：
10.1016/j.biomaterials.2010.02.032
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发表时间：
2010-06-01
期刊：
影响因子：
14
通讯作者：
Beltram, Fabio
中科院分区：
文献类型：
--
作者：
Ferrari, Aldo;Cecchini, Marco;Beltram, Fabio
分化中的神经元和细胞外环境之间的相互作用指导神经系统发育期间细胞极性的建立。发育中的神经元以接触依赖的方式读取局部基底的物理特性，并检索必要的指导线索。在以前的工作中，我们证明了PC12细胞与纳米光栅（亚微米尺寸的脊和槽的交替线）的相互作用促进了双极性和与基底地形的对齐。在这里，我们研究了粘着斑、细胞收缩性和肌动蛋白动力学在这个过程中的作用。利用纳米压印光刻技术和环烯烃共聚物，我们设计的生物相容性纳米结构基板设计的高分辨率活细胞显微镜。我们的研究结果表明，神经元极化和接触指导是基于局部粘连的几何约束，导致其成熟和持久性的角度调制。我们报告ROCK 1/2-肌球蛋白-II通路的活性，并证明ROCK介导的收缩性有助于在神经元分化过程中的极性选择。重要的是，选择过程限制了肌动蛋白支持的膜突起的产生和两极新神经突的形成。建立的极性的维持与NGF刺激无关。总之，我们的结果表明，粘着斑和细胞收缩性稳定地将细胞外环境的拓扑结构与相应的神经元极性状态联系起来（C）2010 Elsevier Ltd保留所有权利。
Interaction between differentiating neurons and the extracellular environment guides the establishment of cell polarity during nervous system development. Developing neurons read the physical properties of the local substrate in a contact-dependent manner and retrieve essential guidance cues In previous works we demonstrated that PC12 cell interaction with nanogratings (alternating lines of ridges and grooves of submicron size) promotes bipolarity and alignment to the substrate topography Here, we investigate the role of focal adhesions, cell contractility, and actin dynamics in this process. Exploiting nanoimprint lithography techniques and a cyclic olefin copolymer, we engineered biocompatible nanostructured substrates designed for high-resolution live-cell microscopy. Our results reveal that neuronal polarization and contact guidance are based on a geometrical constraint of focal adhesions resulting in an angular modulation of their maturation and persistence We report on ROCK1/2-myosin-II pathway activity and demonstrate that ROCK-mediated contractility contributes to polarity selection during neuronal differentiation. Importantly, the selection process confined the generation of actin-supported membrane protrusions and the initiation of new neurites at the poles. Maintenance of the established polarity was independent from NGF stimulation Altogether our results imply that focal adhesions and cell contractility stably link the topographical configuration of the extracellular environment to a corresponding neuronal polarity state (C) 2010 Elsevier Ltd All rights reserved.