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首页> 外文期刊>Cellular and Molecular Bioengineering >Focal Adhesion Induction at the Tip of a Functionalized Nanoelectrode
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Focal Adhesion Induction at the Tip of a Functionalized Nanoelectrode

机译：在功能化的纳米电极的尖端处的局部粘附诱导。

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Cells dynamically interact with their physical micro-environment through the assembly of nascent focal contacts and focal adhesions. The dynamics and mechanics of these contact points are controlled by transmembrane integrins and an array of intracellular adaptor proteins. In order to study the mechanics and dynamics of focal adhesion assembly, we have developed a technique for the timed induction of a nascent focal adhesion. Bovine aortic endothelial cells were approached at the apical surface by a nanoelectrode whose position was controlled with a resolution of 10 s of nanometers using changes in electrode current to monitor distance from the cell surface. Since this probe was functionalized with fibronectin, a focal contact formed at the contact location. Nascent focal adhesion assembly was confirmed using time-lapse confocal fluorescent images of red fluorescent protein—tagged talin, an adapter protein that binds to activated integrins. Binding to the cell was verified by noting a lack of change of electrode current upon retraction of the electrode. This study demonstrates that functionalized nanoelectrodes can enable precisely-timed induction and 3-D mechanical manipulation of focal adhesions and the assay of the detailed molecular kinetics of their assembly.
机译：细胞通过新生的焦点接触和粘着斑的组装动态地与其物理微环境相互作用。这些接触点的动力学和力学由跨膜整合素和一系列细胞内衔接蛋白控制。为了研究粘着斑组装的力学和动力学，我们开发了一种定时诱导新生粘着斑的技术。牛主动脉内皮细胞通过纳米电极到达顶表面，其位置通过电极电流的变化以10 s纳米的分辨率进行控制，以监测距细胞表面的距离。由于该探针已用纤连蛋白功能化，因此在接触部位形成了焦点接触。使用红色荧光蛋白（标记的塔林蛋白，一种与活化的整联蛋白结合的衔接蛋白）的延时共聚焦荧光图像可以证实新生的粘着斑组装。通过注意到在电极缩回时电极电流没有变化来验证与细胞的结合。这项研究表明，功能化的纳米电极可以实现对粘着斑进行精确定时的诱导和3-D机械操作，并可以检测其组装的详细分子动力学。

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