Influence of adhesion and cytoskeletal integrity on fibroblast traction.

Influence of adhesion and cytoskeletal integrity on fibroblast traction.
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粘附和细胞骨架完整性对成纤维细胞牵引的影响。

DOI:
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发表时间:
1996
影响因子:
--
通讯作者:
A. Ott
A. Ott
中科院分区:
--
文献类型:
--
作者:
O. Thoumine;A. Ott

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细胞的伸缩性在细胞形态发生和组织形态形成中起着重要作用。在此背景下,我们研究了细胞牵引力的表达如何依赖于细胞与底物的接触和细胞骨架的组织。对培养在聚二甲基硅氧烷弹性膜上的鸡成纤维细胞的定性观察表明,皱纹模式与肌动蛋白应力纤维和焦点接触的分布有很强的空间关系。为了进一步表征细胞的收缩能力,在底物粘附性、细胞骨架扰动和温度等不同条件下,测量了三磷酸腺苷诱导的成纤维细胞收缩时Triton通透性成纤维细胞的投影面积。在所有条件下,细胞退缩程度与ATP浓度之间的关系均符合酶动力学规律。在明胶包被的基质上培养细胞,降低温度,使用ATP以外的磷酸核苷酸,以及用细胞松弛素D处理细胞,都会降低细胞的回缩率,表明成纤维细胞的牵引力是由温度和ATP依赖的肌动蛋白/肌球蛋白应激纤维滑动机制产生的,并通过焦点粘连传递到底物。诺考达唑或紫杉醇处理细胞不影响三磷酸腺苷刺激时渗透性成纤维细胞的收缩,这表明微管不直接抵抗细胞牵引。用钒酸盐处理细胞增加了细胞的回缩,这表明中间丝有助于传递张力。
Cellular contractility plays an important role in cell morphogenesis and tissue pattern formation. In this context, we examined how the expression of cell traction depends on cell-to-substrate contacts and cytoskeletal organization. Qualitative observation of chick fibroblasts cultured on an elastic film of polydimethylsiloxane indicated a strong spatial relationship between wrinkle pattern and distribution of actin stress fibers and focal contacts. In order to further characterize cell contractility, the projected area of Triton-permeabilized fibroblasts upon ATP-induced retraction was measured in various conditions of substrate adhesivity, cytoskeletal perturbation, and temperature. In all conditions, the relationship between degree of cell retraction and ATP concentration was well described by the laws of enzyme kinetics. Culturing cells on a gelatin-coated substrate, decreasing the temperature, using phosphate ribonucleotides other than ATP, and treating cells with cytochalasin D all diminished the rate of cell retraction, indicating that fibroblast traction is generated by a temperature- and ATP-dependent actin/myosin stress fiber sliding mechanism, transmitted to the substrate through focal adhesions. Treatment of cells with either nocodazole or taxol did not affect retraction of permeabilized fibroblasts upon stimulation with ATP, suggesting that microtubules do not directly resist cell traction. Treatment of cells with vanadate increased cell retraction, suggesting that intermediate filaments help transmit tension.
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