Correlative STED and Atomic Force Microscopy on Live Astrocytes Reveals Plasticity of Cytoskeletal Structure and Membrane Physical Properties during Polarized Migration.
Correlative STED and Atomic Force Microscopy on Live Astrocytes Reveals Plasticity of Cytoskeletal Structure and Membrane Physical Properties during Polarized Migration.
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DOI：
10.3389/fncel.2017.00104
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发表时间：
2017
影响因子：
5.3
通讯作者：
Kaminski CF
中科院分区：
文献类型：
--
作者：
Curry N;Ghézali G;Kaminski Schierle GS;Rouach N;Kaminski CF
细胞骨架结构和膜性质的可塑性对于细胞的极性、黏附和迁移的建立是重要的。在这里，我们提出了一种结合受激发射耗竭(STED)超分辨率成像和原子力显微镜(AFM)的方法来关联活的星形胶质细胞的细胞骨架结构信息和膜物理特性。使用STED兼容染料对细胞骨架进行活细胞成像，同时使用AFM绘制细胞表面拓扑结构，我们获得了星形胶质细胞中高度组织的肌动蛋白和微管网络的前所未有的细节。将来自AFM的机械数据与肌动蛋白和微管蛋白的光学成像相结合，进一步揭示了细胞骨架组织和膜特性之间的联系。使用这种方法，我们说明了划痕诱导的迁移诱导细胞骨架重塑。后者是由星形胶质细胞突起内肌动蛋白和微管成分的极化引起的，这与细胞硬度的变化密切相关。该方法为动态探测活脑细胞的膜结构和功能可塑性开辟了新的途径。它是一种强大的工具，可以为研究生理或病理过程中细胞结构重塑的机制提供新的见解，如脑发育或肿瘤发生。
The plasticity of the cytoskeleton architecture and membrane properties is important for the establishment of cell polarity, adhesion and migration. Here, we present a method which combines stimulated emission depletion (STED) super-resolution imaging and atomic force microscopy (AFM) to correlate cytoskeletal structural information with membrane physical properties in live astrocytes. Using STED compatible dyes for live cell imaging of the cytoskeleton, and simultaneously mapping the cell surface topology with AFM, we obtain unprecedented detail of highly organized networks of actin and microtubules in astrocytes. Combining mechanical data from AFM with optical imaging of actin and tubulin further reveals links between cytoskeleton organization and membrane properties. Using this methodology we illustrate that scratch-induced migration induces cytoskeleton remodeling. The latter is caused by a polarization of actin and microtubule elements within astroglial cell processes, which correlates strongly with changes in cell stiffness. The method opens new avenues for the dynamic probing of the membrane structural and functional plasticity of living brain cells. It is a powerful tool for providing new insights into mechanisms of cell structural remodeling during physiological or pathological processes, such as brain development or tumorigenesis.