利用免疫染色定量x射线和热诱导的正常细胞和转化细胞中的多个微管组织中心。

Basic and applied histochemistry Pub Date : 1988-01-01
A U Lobreau, G P Raaphorst, J G Szekely
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引用次数: 0

摘要

用微管蛋白抗体间接免疫荧光法观察对照、辐照和加热C3H 10T1/2小鼠胚胎细胞和辐照转化亚群R1和R25中的微管组织中心(MTOC)。微管用诺可达唑解聚后,在新鲜培养基中孵育5分钟重组MTOC。R1系的MTOC/cell分布与亲本10T1/2系或R25的MTOC/cell分布相似,但R1系的MTOC/cell分布不同。辐照后,正常细胞和辐射转化细胞在10 Gy照射下孵育24或48 h后出现多个MTOC。辐照后细胞微管重构的多发灶表明,辐射损伤表现在细胞质结构元件上。三种细胞系热处理(43℃93 min和45℃25 min)后,MTOC被破坏,许多细胞在24或48 h时没有明显的组织中心,而另一些细胞则有大量小的微管重组中心。处理后R25细胞中MTOC/cell的分布与10T1/2细胞系的分布相似,而与另一辐射转化细胞系R1的分布不同。因此,在MTOC中看到的辐射或热响应不依赖于细胞转化。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
The use of immunostaining to quantify x-ray and heat-induced multiple microtubule organizing centers in normal and transformed cells.

Microtubule organizing centers (MTOC) in control, irradiated and heated C3H 10T1/2 mouse embryo cells and two radiation-transformed sublines, R1 and R25, were made visible by indirect immunofluorescence using antibody against tubulin. The MTOC were reformed by 5-min incubation in fresh medium after the microtubules were depolymerized with nocodazole. The R1 line had a different distribution of MTOC/cell than the parent 10T1/2 line or R25, which had similar distributions. After irradiation, multiple MTOC appeared in the normal and radiation-transformed cells irradiated to 10 Gy and incubated for 24 or 48 h. The multiple foci of microtubule reformation in the irradiated cells indicate that radiation damage is expressed in structural elements in the cytoplasm. After heat treatment of the three cell lines (43 degrees C for 93 min and 45 degrees C for 25 min), the MTOC were disrupted and many cells did not have visible organizing centers at 24 or 48 h, while others had a large number of small centers of microtubule reformation. The distribution of MTOC/cell seen in R25 cells after the treatment had similar patterns to those of the 10T1/2 line rather than to those of the other radiation-transformed line, R1. Thus, the radiation or heat response seen in the MTOC is not dependent upon cell transformation.

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