STRUCTURE OF THE MITOTIC APPARATUS AND CHROMOSOMES AFTER HYPOTONIC TREATMENT OF MAMMALIAN-CELLS INVITRO
STRUCTURE OF THE MITOTIC APPARATUS AND CHROMOSOMES AFTER HYPOTONIC TREATMENT OF MAMMALIAN-CELLS INVITRO
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DOI：
10.1159/000131438
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发表时间：
1980-01-01
期刊：
影响因子：
--
通讯作者：
PEPPER, DA
中科院分区：
文献类型：
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作者：
BRINKLEY, BR;COX, SM;PEPPER, DA
将培养的哺乳动物细胞[中国仓鼠成纤维细胞、瑞士小鼠3T3成纤维细胞和三趾鼠肾PtK2细胞]短暂暴露于加有不同稀释度蒸馏水的低渗培养液中。对短暂暴露于低渗条件下的有丝分裂细胞进行了电子显微镜观察和微管蛋白免疫荧光检测。类似的观察也发生在暴露在低渗介质中然后又回到等渗条件下的细胞上。对有丝分裂器和染色体的结构和物理性质有了深入的了解。纺锤体微管通过低渗处理可逆地解聚。低渗15分钟后，纺锤体纤维消失，只剩下2个明亮的纺锤体极。当低张肿胀的细胞在固定前返回等渗液15min时，纺锤体重新组装。在从低张治疗中恢复的细胞中发现了几种类型的纺锤体异常。低渗处理使染色体大大扩张，动粒被打乱。当处理的细胞回到等渗培养基时，染色体重新凝聚，动粒重新出现。短暂的低渗处理对细胞活力没有不良影响，在研究有丝分裂器的结构和物理性质时可能会被证明是有用的。
Cultured mammalian cells [Chinese hamster fibroblasts, Swiss mouse 3T3 fibroblasts and Potorous tridactylus kidney PtK2 cells] were exposed briefly to culture media made hypotonic with various dilutions of distilled water. EM observations and tubulin immunofluorescence were carried out on mitotic cells that had been briefly exposed to hypotonic conditions. Similar observations were made on cells that had been exposed to hypotonic medium and then returned to isotonic conditions. Insight into the structure and physical properties of the mitotic apparatus and chromosomes was gained. Spindle microtubules were reversibly depolymerized by hypotonic treatments. Spindle fibers disappeared after a 15 min exposure to hypotonic medium, leaving only 2 brightly fluorescent spindle poles. When hypotonically swollen cells were returned to isotonic medium for 15 min prior to fixation, the spindle was reassembled. Several types of spindle aberrations were noted in cells recovering from hypotonic treatment. Chromosomes were greatly expanded by hypotonic treatment, and the kinetochores were disrupted. When treated cells were returned to isotonic culture medium, the chromosomes recondensed and the kinetochores reappeared. Brief hypotonic treatments had no adverse effect on cell viability and may prove useful in investigations of the structure and physical properties of the mitotic apparatus.