Mechanical dynamics of focal adhesions of vascular endothelial cells using nano-imaging

使用纳米成像研究血管内皮细胞粘着斑的机械动力学

• 批准号: 15086203
• 负责人: SATO Masaaki
• 金额: $ 22.21万
• 依托单位: Tohoku University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research on Priority Areas
• 财政年份: 2003
• 资助国家: 日本
• 起止时间: 2003 至 2006
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15086203/
• 关键词: Focal adhesions; Actin filaments; Gene transfection; Flow exposure; Stretching; Micromachining; Mechanotransduction; 細胞骨格; 血管内皮細胞; 張力負荷; せん断応力負荷; 静水圧負荷; マイクロポスト; せん断応力; 微小管; 葉状仮足; バイオメカニクス; 内皮細胞; 焦点接着班; 近接場光

The goal of this project is to study dynamics of FAT domain in focal adhesions and associated actin filaments in order to understand remodeling process of adherent cells in response to external mechanical stimuli. In particular, newly developed experimental techniques here include a time-course observation of dynamics of FAT domain and actin filaments under application of fluid flow and a stretching system using an elastic membrane with nonuniform surface structure. First, we have developed an experimental system which enables to observe dynamics of cells using RFP-conjugated FAT-domain vector and GFP-conjugated actin vector. The result showed that actin filament structure was reconfigured in a close association with appearance/disappearance of FAT domain. Second, PDMS membrane with an array of micropillars has been fabricated by using soft lithography technique to locally apply stretching between focal adhesions. Endothelial cells formed focal adhesion complexes at peripheries of the top of the micropillars and developed stress fibers, thick actin bundles, perpendicular to the direction of stretch. Finally, we have developed a micro-tensile tester to measure mechanical properties of stress fibers, which would be critical for understanding of intracellular stress transfer. Tensile tests were, for the first time, carried out on stress fibers chemically isolated from endothelial cells, providing a load-displacement relationship. It was found that the load-displacement curve was nonlinear with high extensibility. We have successfully studied important roles of cytoskeletons and focal adhesions of mechanically stimulated cells in intracellular force transmission mechanism.

本项目旨在研究局灶性黏附中FAT结构域及相关肌动蛋白丝的动力学，以了解黏附细胞在外部机械刺激下的重塑过程。特别是，新开发的实验技术包括在流体流动作用下的FAT结构域和肌动蛋白丝动力学的时间过程观察和使用具有非均匀表面结构的弹性膜的拉伸系统。首先，我们开发了一个实验系统，可以使用rfp共轭的脂肪结构域载体和gfp共轭的肌动蛋白载体来观察细胞的动力学。结果表明，肌动蛋白丝结构的重构与FAT结构域的出现/消失密切相关。其次，利用软光刻技术在黏着点之间局部施加拉伸，制备了具有微柱阵列的PDMS膜。内皮细胞在微柱顶部周周形成局灶性黏附复合物，并形成应力纤维，粗肌动蛋白束，垂直于拉伸方向。最后，我们开发了一种微拉伸测试仪来测量应力纤维的机械性能，这对于理解细胞内应力传递至关重要。拉伸试验首次对从内皮细胞中化学分离出来的应力纤维进行了拉伸试验，提供了载荷-位移关系。结果表明，荷载-位移曲线是非线性的，具有较高的可拓性。我们成功地研究了细胞骨架和机械刺激细胞的局灶黏附在细胞内力传递机制中的重要作用。

项目成果

Endothelial Biomechanics : Focusing on the Dynamic Behavior of Cytoskeletal Structure

内皮生物力学：关注细胞骨架结构的动态行为

• 发表时间: 2006
• 作者: Hiroshi Wada (editor);Ryo Sudo;Y.Tanimoto;Masaaki Sato
• 通讯作者: Masaaki Sato

Biomechanics at Micro- and Nanoscale levels. Vol. I

微米和纳米级的生物力学。

• 发表时间: 2005
• 作者: Iguchi.H;Sakai.J;et al.;Hiroshi Wada
• 通讯作者: Hiroshi Wada

Strain rate-dependent tensile properties of single stress fibers isolated from smooth muscle cells

从平滑肌细胞中分离出的单应力纤维的应变率依赖性拉伸特性

• 发表时间: 2007
• 作者: T.Nakagawa;T.Yokoyama;村上輝夫(編著);S.Kohtani;Tsubasa Matsui
• 通讯作者: Tsubasa Matsui

マトリックスデバイスを用いた平滑筋細胞の形態制御と牽引力測定

使用矩阵装置平滑肌细胞的形态控制和牵引力测量

• 发表时间: 2006
• 作者: S.S.Staniland;W.Fujita;Y.Umezono;K.Awaga K;P.J.Camp;S.J.Clark;N.Robertson;Y Tanimoto;山田宏;Hiroyuki Asami;Wada H;大橋俊朗
• 通讯作者: 大橋俊朗

平滑筋細胞の初期ひずみの計測

平滑肌细胞初始应变的测量

• 发表时间: 2004
• 作者: M.Sakai et al.;出口真次
• 通讯作者: 出口真次