Role of the F-actin cytoskeleton in the RVD and RVI processes in Ehrlich ascites tumor cells
Role of the F-actin cytoskeleton in the RVD and RVI processes in Ehrlich ascites tumor cells
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DOI：
10.1006/excr.1999.4615
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发表时间：
1999-10-10
影响因子：
3.7
通讯作者：
Hoffmann, EK
中科院分区：
文献类型：
--
作者：
Pedersen, SF;Mills, JW;Hoffmann, EK
在埃利希腹水肿瘤细胞中，使用定量罗丹明-phalloidin测定、共聚焦激光扫描显微镜和电子细胞定尺研究了f -肌动蛋白细胞骨架在细胞体积调节中的作用。与等渗（310 mOsm）对照细胞相比，低渗刺激（180 mOsm）与1和3分钟时细胞f -肌动蛋白含量下降有关，高渗刺激（600 mOsm）与1、3和5分钟时细胞f -肌动蛋白含量增加有关。在固定完整的埃利希细胞中共聚焦显示，渗透挑战主要影响细胞皮层区域的f -肌动蛋白，而其他细胞区域的f -肌动蛋白未见明显变化。使用0.5 μ M的细胞松弛素B （CB）、细胞松弛素D （CD）或毛球蛋白C （ChtC）研究了f -肌动蛋白细胞骨架在RVD中的可能作用。毛球蛋白C是一种与f -肌动蛋白亲和力很小或没有亲和力的细胞松弛素类似物。与渗透性肿胀的对照细胞相比，用0.5 μ M CB预处理3分钟的细胞在低渗肿胀后体积恢复较慢，而CD和chtc处理的细胞则没有。此外，cb处理的细胞肿胀后的最大细胞体积减少，而CD或chtc处理的细胞则没有。在使用RVD/RVI方案施加高渗刺激后，cb处理的细胞从收缩中恢复较慢，而CD或chtc处理的细胞则没有，而收缩后的最小细胞体积在这两种处理中都没有改变。结果表明，渗透细胞的肿胀和收缩分别引起埃利希细胞中f -肌动蛋白含量的减少和增加。0.5 μ M CB对RVD和RVI过程有抑制作用，而0.5 μ M CD对肌动蛋白的特异性更强。(C) 1999学术出版社。
The role of the F-actin cytoskeleton in cell volume regulation was studied in Ehrlich ascites tumor cells, using a quantitative rhodamine-phalloidin assay, confocal laser scanning microscopy, and electronic cell sizing. A hypotonic challenge (180 mOsm) was associated with a decrease in cellular F-actin content at 1 and 3 min and a hypertonic challenge (600 mOsm) with an increase in cellular F-actin content at 1, 3, and 5 min, respectively, compared to isotonic (310 mOsm) control cells. Confocal visualization of F-actin in fixed, intact Ehrlich cells demonstrated that osmotic challenges mainly affect the F-actin in the cortical region of the cells, with no visible changes in F-actin in other cell regions. The possible role of the F-actin cytoskeleton in RVD was studied using 0.5 mu M cytochalasin B (CB), cytochalasin D (CD), or chaetoglobosin C (ChtC), a cytochalasin analog with little or no affinity for F-actin.Recovery of cell volume after hypotonic swelling was slower in cells pretreated for 3 min with 0.5 mu M CB, but not in CD- and ChtC-treated cells, compared to osmotically swollen control cells. Moreover, the maximal cell volume after swelling was decreased in CB-treated, but not in CD- or Chtc-treated cells. Following a hypertonic challenge imposed using the RVD/RVI protocol, recovery from cell shrinkage was slower in CB-treated, but not in CD- or Chtc-treated cells, whereas the minimal cell volume after shrinkage was unaltered by either of these treatments. It is concluded that osmotic cell swelling and shrinkage elicit a decrease and an increase in the F-actin content in Ehrlich cells, respectively. The RVD and RVI processes are inhibited by 0.5 mu M CB, but not by 0.5 mu M CD, which is more specific for actin. (C) 1999 Academic Press.