Indirect immunofluorescence of microtubules in Dictyostelium discoideum. A study with polyclonal and monoclonal antibodies to tubulins.
Indirect immunofluorescence of microtubules in Dictyostelium discoideum. A study with polyclonal and monoclonal antibodies to tubulins.
复制标题
盘基网柄菌微管的间接免疫荧光。
DOI：
10.1016/0014-4827(84)90367-7
复制
发表时间：
1984
影响因子：
3.7
通讯作者：
J. D. De Mey
中科院分区：
文献类型：
--
作者：
U. Roos;M. De Brabander;J. D. De Mey
阿米巴德。用戊二醛原位固定盘状软骨覆盖片，用Triton X-100渗透。在测试的六种抗体中，只有一种酵母微管蛋白的单抗持续发出明亮的荧光。DAPI复染有助于识别间期和有丝分裂阶段。间期阿米巴中的大多数微管(MT)来自一个核相关的中心，该中心具有一个非荧光核心。有丝分裂早期的阿米巴几乎完全缺乏细胞质MT。初生期的纺锤体为明亮的荧光点，而早中期的纺锤体为短杆。中期和后期核的纺锤体较长，有的由多条荧光线组成。末期和后期纺锤体上的星体MTs较多。在有丝分裂后细胞中，ASTs明显转化为MTS的间期复合体，而中心纺锤体的杆状残余消失。与细胞分裂相关的MT系统的周期性变化与高等真核生物中观察到的相似，可能反映了阿米巴运动行为的变化，而不是细胞形状的变化。
Amebae ofD. discoideumon coverslips were fixed in situ with glutaraldehyde and permeabilized with Triton X-100. Of six antibodies tested, only a monoclonal antibody to yeast tubulin consistently gave bright fluorescence. Counterstaining with DAPI facilitated the identification of interphase and mitotic stages. Most microtubules (MTs) in interphase amebae emanated from a nucleus-associated centre that had a non-fluorescent core. Amebae in early stages of mitosis lacked cytoplasmic MTs almost entirely. The nascent spindle in prophase appeared as a brightly fluorescent dot, whereas the prometaphase spindle was a short rod. Spindles in metaphase and anaphase nuclei were more elongate, some consisting of several fluorescent lines. Astral MTs were prominent on spindles in anaphase and telophase. Asters are obviously converted to the interphase complex of MTs in post-mitotic cells, while the shaft-like remnant of the central spindle disappears. The cyclical changes in the MT system related to cell division resemble those observed in higher eukaryotes and probably reflect changes in the locomotory behavior of the amebae rather than changes in cell shape.