Hydrostatic pressure has different effects on the assembly of tubulin, actin, myosin II, vinculin, talin, vimentin, and cytokeratin in mammalian tissue cells
Hydrostatic pressure has different effects on the assembly of tubulin, actin, myosin II, vinculin, talin, vimentin, and cytokeratin in mammalian tissue cells
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DOI：
10.1006/excr.1996.0278
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发表时间：
1996-09-15
影响因子：
3.7
通讯作者：
Salmon, ED
中科院分区：
文献类型：
--
作者：
Crenshaw, HC;Allen, JA;Salmon, ED
已知数百个大气压范围内的静水压力会破坏组织培养细胞中的细胞骨架组织，使细胞形状发生深刻变化。这些作用的分子机制尚不清楚。为了确定压力对细胞骨架的影响，从而提供更好的分子机制指标，我们使用荧光抗体染色比较了HeLa细胞和大鼠骨肉瘤细胞（ROS-17/2.8）在400 atm不同压力下的7种不同细胞骨架蛋白的组织。300atm或更高的压力导致两系细胞“聚集”并撤回其片层延伸。然而，这种反应在细胞群体中是不同的，一些细胞在压力下继续扩散，导致它们的邻居聚集起来。最能抵抗舍入的是那些与其他细胞接触的细胞，以及偶尔出现的巨细胞。正如预期的那样，圆形细胞显示肌动蛋白应激纤维和血管蛋白和talin在局灶接触处的破坏。非圆形细胞在这些相同蛋白质的组织中表现出较少的破坏，微管和肌球蛋白II丝在两种细胞类型中，无论是圆形还是非圆形，都表现出对400 atm压力的抵抗力。然而，在HeLa细胞中，当压力超过200 atm时，中间丝，静脉蛋白和细胞角蛋白解聚并形成小泡，这种情况发生在圆形和非圆形细胞中。在没有细胞角蛋白的骨肉瘤细胞中，波形蛋白不能解聚。我们讨论了可能解释这些压力反应的不同机制，包括对蛋白质聚合平衡的直接影响和对控制细胞骨架组织的调节机制（如磷酸化途径）的不太直接的影响。后一种解释似乎更符合细胞群体内反应的可变性，以及一种细胞系与另一种细胞系之间vimentin反应的差异。(C) 1996学术出版社，Inc.
Hydrostatic pressures in the range of hundreds of atmospheres are known to disrupt cytoskeletal organization in tissue culture cells, with profound changes in cell shape. The molecular mechanisms of these effects are poorly understood. To determine the effect of pressure on the cytoskeleton, and thus to provide better indicators of the molecular mechanisms, we used fluorescent antibody staining to compare the organizations of seven different cytoskeletal proteins in HeLa cells and rat osteosarcoma cells (ROS-17/2.8) subjected to different pressures up to 400 atm. Pressures of 300 atm or more caused cells of both lines to ''round up'' and to withdraw their lamellar extensions. However, this response varied within a population of cells, with some cells remaining spread at pressures that caused their neighbors to round up. The most resistant to rounding were those cells touching other cells, and the occasional giant cells. As expected, the rounded cells showed disruption of actin stress fibers and of vinculin and talin at focal contacts. The unrounded cells showed less disruption in the organization of these same proteins, Microtubules and myosin II filaments appeared resistant to 400 atm pressure in both cell types, whether rounded or unrounded. However, in HeLa cells, the intermediate filaments, vimentin and cytokeratin, depolymerized and formed small vesicles when pressures exceeded 200 atm, and this occurred in rounded as well as unrounded cells. In osteosarcoma cells, which do not have cytokeratin, vimentin did not depolymerize. We discuss different mechanisms that might explain these responses to pressure, including direct effects on the equilibria of protein polymerization and less direct effects on regulatory mechanisms, such as phosphorylation pathways, that control cytoskeletal organization. The latter type of explanation seems more consistent with both the variability of response within cell populations and the difference in vimentin's response in one cell line compared with the other. (C) 1996 Academic Press, Inc.