Coss R A, Pickett-Heaps J D
J Cell Biol. 1974 Oct;63(1):84-98. doi: 10.1083/jcb.63.1.84.
Cell division in vegetative filaments of the green alga Oedogonium cardiacum is presented as an experimental system. We report on how we have used this system to study the effects of isopropyl N-phenylcarbamate (IPC) on the mitotic apparatus and on the phycoplast, a planar array of cytokinetic microtubules. Polymerization of microtubules was prevented when filaments, synchronized by a light/dark regime and chilled (2 degrees C) while in metaphase or just before phycoplast formation, were exposed to 5.5 x 10(-4) M IPC and then returned to room temperature. Spindles reformed or phycoplasts formed when these filaments were transferred to growth medium free of IPC. However, the orientation of both microtubular systems was disturbed: the mitotic apparatus often contained three poles, frequently forming three daughter nuclei upon karyokinesis; the phycoplast was often stellate rather than planar, and it sometimes was displaced to the side of both daughter nuclei, resulting in a binucleate and an anucleate cell upon cytokinesis. Our results suggest that IPC (a) prevents the assembly of microtubules, (b) increases the number of functional polar bodies, and (c) affects the orientation of microtubules in O. cardiacum. High voltage (1,000 kV) electron microscopy of 0.5-microm thick sections allowed us to visualize the polar structures, which were not discernible in thin sections.
绿藻心形鞘藻营养丝中的细胞分裂被作为一个实验系统呈现出来。我们报告了如何利用这个系统来研究异丙基-N-苯基氨基甲酸酯(IPC)对有丝分裂器和藻质体(一种细胞分裂微管的平面阵列)的影响。当通过光/暗周期同步并在中期或即将形成藻质体时进行冷却(2摄氏度)的藻丝暴露于5.5×10⁻⁴ M IPC,然后恢复到室温时,微管的聚合被阻止。当这些藻丝转移到不含IPC的生长培养基中时,纺锤体重新形成或藻质体形成。然而,两个微管系统的取向都受到了干扰:有丝分裂器通常包含三个极,在核分裂时经常形成三个子核;藻质体通常呈星状而非平面状,有时会移位到两个子核的一侧,导致在胞质分裂时形成一个双核细胞和一个无核细胞。我们的结果表明,IPC(a)阻止微管的组装,(b)增加功能性极体的数量,并且(c)影响心形鞘藻中微管的取向。对0.5微米厚切片进行的高电压(1000 kV)电子显微镜观察使我们能够可视化在薄切片中无法辨别的极性结构。