Probing in vivo dynamics of mitochondria and cortical actin networks using high‐speed atomic force/fluorescence microscopy
Probing in vivo dynamics of mitochondria and cortical actin networks using high‐speed atomic force/fluorescence microscopy
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DOI：
10.1111/gtc.12204
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发表时间：
2015-02
期刊：
影响因子：
2.1
通讯作者：
A. Yoshida;Nobuaki Sakai;Yoshitsugu Uekusa;Katashi Deguchi;Jamie L. Gilmore;Masahiro Kumeta;Shuichi Ito;K. Takeyasu
中科院分区：
文献类型：
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作者：
A. Yoshida;Nobuaki Sakai;Yoshitsugu Uekusa;Katashi Deguchi;Jamie L. Gilmore;Masahiro Kumeta;Shuichi Ito;K. Takeyasu
细胞膜和膜下结构的动力学紧密相连，促进着细胞的各种活动。虽然细胞表面研究和皮质肌动蛋白研究已经显示了细胞膜和肌动蛋白网络的独立机制，但很难全面了解活细胞中这些结构的动力学。在这里，我们使用原子力/光学显微镜组合系统在纳米尺度分辨率下分析活细胞中基于膜的细胞事件。COS‐7细胞表面成像显示了与内吞作用和胞吐作用相对应的膜内陷事件的详细结构特性。此外，线粒体的运动和皮层F - actin网络的时空动态在体内被直接可视化。大小在1.7 × 104 ~ 1.4 × 105 nm2之间的皮质肌动蛋白微结构域被新出现的肌动蛋白丝动态重排，有时伴有膜内翻，表明这些事件与肌动蛋白周转维持的膜下组织的动态调节有关。这些结果为整个细胞膜机制的结构方面提供了新的见解，可以用高时间和空间分辨率进行可视化。
The dynamics of the cell membrane and submembrane structures are closely linked, facilitating various cellular activities. Although cell surface research and cortical actin studies have shown independent mechanisms for the cell membrane and the actin network, it has been difficult to obtain a comprehensive understanding of the dynamics of these structures in live cells. Here, we used a combined atomic force/optical microscope system to analyze membrane‐based cellular events at nanometer‐scale resolution in live cells. Imaging the COS‐7 cell surface showed detailed structural properties of membrane invagination events corresponding to endocytosis and exocytosis. In addition, the movement of mitochondria and the spatiotemporal dynamics of the cortical F‐actin network were directly visualized in vivo. Cortical actin microdomains with sizes ranging from 1.7 × 104 to 1.4 × 105 nm2 were dynamically rearranged by newly appearing actin filaments, which sometimes accompanied membrane invaginations, suggesting that these events are integrated with the dynamic regulation of submembrane organizations maintained by actin turnovers. These results provide novel insights into the structural aspects of the entire cell membrane machinery which can be visualized with high temporal and spatial resolution.