Gambino J, Bergen L G, Morris N R
J Cell Biol. 1984 Sep;99(3):830-8. doi: 10.1083/jcb.99.3.830.
We used immunofluorescent microscopy to characterize microtubule (MT) architecture in wild-type and mutant protoplasts of Aspergillus nidulans at interphase and at mitosis. Because the visualization of MTs by immunofluorescence is technically difficult in intact hyphae of A. nidulans, we developed a method for removing the cell wall under conditions that do not perturb cell physiology, as evidenced by the fact that the resulting protoplasts undergo nuclear division at a normal rate and that cell cycle mutant phenotypes are expressed at restrictive temperature. Interphase cells exhibited an extensive network of cytoplasmic MTs. During mitosis the cytoplasmic MTs mostly disappeared and an intranuclear mitotic spindle appeared. We have previously shown that the benA 33 beta-tubulin mutation causes hyperstabilization of the mitotic spindle, and we have presented additional indirect evidence that suggested that the tubA1 and tubA4 alpha-tubulin mutations destabilize spindle MTs. In this paper, we show that the benA33 mutation increases the stability of cytoplasmic MTs as well as spindle MTs and that the tubA1 and tubA4 mutations destabilize both spindle and cytoplasmic MTs.
我们利用免疫荧光显微镜对构巢曲霉野生型和突变型原生质体在间期和有丝分裂期的微管（MT）结构进行了表征。由于在构巢曲霉完整的菌丝中通过免疫荧光观察微管在技术上存在困难，我们开发了一种在不干扰细胞生理的条件下去除细胞壁的方法，这一点可由以下事实证明：所得原生质体以正常速率进行核分裂，并且细胞周期突变体表型在限制温度下得以表达。间期细胞呈现出广泛的细胞质微管网。在有丝分裂期间，细胞质微管大多消失，并且出现了核内有丝分裂纺锤体。我们之前已经表明，benA 33β-微管蛋白突变会导致有丝分裂纺锤体超稳定，并且我们还提供了额外的间接证据表明tubA1和tubA4α-微管蛋白突变会使纺锤体微管不稳定。在本文中，我们表明benA33突变增加了细胞质微管以及纺锤体微管的稳定性，而tubA1和tubA4突变则使纺锤体和细胞质微管都不稳定。
