Knowles R, LeClerc N, Kosik K S
Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston.
Cell Motil Cytoskeleton. 1994;28(3):256-64. doi: 10.1002/cm.970280308.
Insect Sf9 cells usually elaborate a highly characteristic single process when infected with a baculovirus encoding recombinant human tau. The processes are unbranched, of uniform caliber, and contain bundles of microtubules. Because taxol treatment alone does not induce process outgrowth in these cells, it is believed that tau confers properties on microtubules that permits the conversion of microtubule assembly into the formation of processes. Here we have analyzed the reorganization of both actin filaments and microtubules during process initiation. A zone of organelle exclusion representing the focal reorganization of actin at one pole of the cell anticipated process emergence. A relationship between actin organization and process emergence was also suggested by a shift from single to multiple process formation after treatment with cytochalasin D. The rate of process elongation doubled after cytochalasin treatment of tau-expressing cells. The increase in rate was due to the inhibition of the growth arrest phases which occur in the absence of cytochalasin. In contrast, Sf9 cells treated with cytochalasin after more than 20 h of tau expression were relatively resistant to the drug's effects. We conclude that actin and microtubules are specifically reorganized during tau-induced process outgrowth and that a dynamic relationship between actin filaments and microtubules effects process formation.
昆虫Sf9细胞在感染编码重组人tau蛋白的杆状病毒时,通常会形成一个高度特征性的单一突起。这些突起没有分支,管径均匀,并且含有微管束。由于单独使用紫杉醇处理不会诱导这些细胞长出突起,因此人们认为tau蛋白赋予微管一些特性,使得微管组装能够转化为突起的形成。在这里,我们分析了突起形成过程中肌动蛋白丝和微管的重组情况。一个代表细胞一极肌动蛋白局部重组的细胞器排除区先于突起出现。用细胞松弛素D处理后,细胞从单个突起形成转变为多个突起形成,这也表明了肌动蛋白组织与突起出现之间的关系。在用细胞松弛素处理表达tau蛋白的细胞后,突起伸长速率增加了一倍。速率的增加是由于抑制了在没有细胞松弛素的情况下出现的生长停滞阶段。相反,在tau蛋白表达超过20小时后用细胞松弛素处理的Sf9细胞对该药物的作用相对耐药。我们得出结论,在tau蛋白诱导的突起生长过程中,肌动蛋白和微管会发生特异性重组,并且肌动蛋白丝和微管之间的动态关系影响突起的形成。