Rosenbaum J L, Carlson K
J Cell Biol. 1969 Feb;40(2):415-25. doi: 10.1083/jcb.40.2.415.
The cilia of Tetrahymena were amputated by the use of a procedure in which the cells remained viable and regenerated cilia. Deciliated cells were nonmotile, and cilia regeneration was assessed by scoring the percentage of motile cells at intervals following deciliation. After a 30-min lag, the deciliated cells rapidly recovered motility until more than 90% of the cells were motile at 70 min after amputation. Cycloheximide inhibited both protein synthesis and cilia regeneration. This indicated that cilia formation in Tetrahymena was dependent on protein synthesis after amputation. Conversely, colchicine was found to inhibit cilia regeneration without affecting either RNA or protein synthesis. This observation suggested the action of colchicine to be an interference with the assembly of ciliary subunit proteins. The finding that colchicine binds to microtubule protein subunits isolated from cilia and flagella (13) supports this possibility. The potential of the colchicine-blocked cilia-regenerating system in Tetrahymena for studying the assembly of microtubule protein subunits during cilia formation and for isolating ciliary precursor proteins is discussed.
通过一种能使细胞保持存活并可再生纤毛的程序,切除了四膜虫的纤毛。去纤毛的细胞无法运动,通过在去纤毛后的不同时间间隔对能运动细胞的百分比进行评分来评估纤毛再生情况。经过30分钟的延迟后,去纤毛的细胞迅速恢复运动能力,到截肢后70分钟时,超过90%的细胞能够运动。环己酰亚胺抑制蛋白质合成和纤毛再生。这表明四膜虫截肢后纤毛的形成依赖于蛋白质合成。相反,发现秋水仙碱抑制纤毛再生,而不影响RNA或蛋白质合成。这一观察结果表明秋水仙碱的作用是干扰纤毛亚基蛋白的组装。秋水仙碱与从纤毛和鞭毛中分离出的微管蛋白亚基结合这一发现支持了这种可能性。讨论了四膜虫中被秋水仙碱阻断的纤毛再生系统在研究纤毛形成过程中微管蛋白亚基组装以及分离纤毛前体蛋白方面的潜力。