Brinkley B R, Fuller E M, Highfield D P
Proc Natl Acad Sci U S A. 1975 Dec;72(12):4981-5. doi: 10.1073/pnas.72.12.4981.
Monospecific antibody directed against bovine brain tubulin was used as an immunofluorescent probe to evaluate the distribution of microtubules in normal and transformed cells grown in tissue culture. The fluorescent staining pattern of transformed and nontransformed cells is significantly different and may be used in conjunction with other morphological features to identify transformants in mixed cell populations. Normal cells are flattened, elongated, and fibroblastic; they display numerous Colcemid-sensitive fluorescent cytoplasmic filaments, presumably microtubules. Transformed cells, however, are smaller, more polygonal in shape, and contain very few cytoplasmic tubules. During mistosis the cytoplasmic microtubule complex of normal cells completely disappears, but reappears after cell division. Treatment of transformed cells with dibutyry-adenosine 3':5'-cyclic monophosphate plus testosterone or theophylline restores the normal fibroblastic appearance of the cells and stimulates the assembly of numerous cytoplasmic microtubules. This study provides further evidence for two separate microtubule entities in cycling nontransformed cells: a cytoplasmic microtubule complex and the microtubules of the mitotic spindle. Although an interchange of tubulin dimers seems to exist between microtubules in the two systems, control of tubule assembly may be under separate constraints. Stimulation of cytoplasmic microtuble assembly in transformed cells by derivatives of adenosine 3':5'-cycle monophosphate suggests that impairment of the cytoplasmic microtubule complex in these cells may be due to suboptimal levels of adenosine 3':5'-cyclic monophosphate.
用针对牛脑微管蛋白的单特异性抗体作为免疫荧光探针，来评估在组织培养中生长的正常细胞和转化细胞中微管的分布。转化细胞和未转化细胞的荧光染色模式有显著差异，可与其他形态学特征一起用于识别混合细胞群体中的转化体。正常细胞扁平、细长，呈成纤维细胞形态；它们显示出许多对秋水仙酰胺敏感的荧光细胞质细丝，推测为微管。然而，转化细胞较小，形状更呈多边形，且细胞质微管极少。在有丝分裂期间，正常细胞的细胞质微管复合体完全消失，但在细胞分裂后重新出现。用二丁酰腺苷3':5'-环磷酸酯加睾酮或茶碱处理转化细胞，可使细胞恢复正常的成纤维细胞外观，并刺激大量细胞质微管的组装。这项研究为处于细胞周期的未转化细胞中两个独立的微管实体提供了进一步的证据：细胞质微管复合体和有丝分裂纺锤体的微管。尽管两个系统中的微管之间似乎存在微管蛋白二聚体的交换，但微管组装的控制可能受不同的限制。腺苷3':5'-环磷酸酯衍生物对转化细胞中细胞质微管组装的刺激表明，这些细胞中细胞质微管复合体的损伤可能是由于腺苷3':5'-环磷酸酯水平不足所致。
