Fujiu K, Numata O
Institute of Biological Sciences, University of Tsukuba, Tsukuba, Ibaraki, Japan.
Cell Motil Cytoskeleton. 2000 May;46(1):17-27. doi: 10.1002/(SICI)1097-0169(200005)46:1<17::AID-CM3>3.0.CO;2-C.
We developed a modified immunofluorescence protocol that permitted visualization of microtubules inside the macronucleus of the ciliate Tetrahymena. Although the amitotically dividing macronucleus lacks a spindle, an elaborate system of microtubules is assembled inside the macronucleus and between the macronucleus and the cortex. Microtubules could not be detected inside the interphase macronuclei. The early stage of macronuclear division was associated with the assembly of short macronuclear microtubules that localized randomly. The intramacronuclear microtubules were subsequently organized in a radial manner. During elongation of the macronucleus, the distribution of macronuclear microtubules changed from radial to parallel. During constriction of the macronucleus, dense and tangled macronuclear microtubules were detected at the region of nuclear constriction. In the cytosol, microtubules were linking the macronucleus and cell cortex. During recovery after drug-induced depolymerization, microtubules reassembled at multiple foci inside the macronucleus in close proximity to the chromatin. We propose that these microtubules play roles in chromatin partitioning, macronuclear constriction, and positioning of the macronucleus in relation to the cell cortex.
我们开发了一种改良的免疫荧光方案,可实现对纤毛虫四膜虫大核内微管的可视化观察。尽管进行无丝分裂的大核缺乏纺锤体,但在大核内部以及大核与皮层之间会组装一个复杂的微管系统。在间期大核内未检测到微管。大核分裂的早期与短的大核微管组装有关,这些微管随机定位。随后,大核内的微管呈放射状排列。在大核伸长过程中,大核微管的分布从放射状变为平行状。在大核缢缩过程中,在核缢缩区域检测到密集且缠结的大核微管。在细胞质中,微管将大核与细胞皮层相连。在药物诱导解聚后的恢复过程中,微管在大核内多个靠近染色质的位点重新组装。我们认为这些微管在染色质分配、大核缢缩以及大核相对于细胞皮层的定位中发挥作用。