Furcht L T, Scott R E
Biochim Biophys Acta. 1975 Aug 20;401(2):213-20. doi: 10.1016/0005-2736(75)90305-3.
The intrinsic organization of the plasma membrane differs in normal and transformed cells. With the technique of freeze fracture and electron microscopy contact inhibited 3T3 cells have been shown to contain aggregated plasma membrane intramembranous particles, while transformed cells demonstrate a uniform particle distribution. The distribution of intramembrous particles in transformed cells can be affected by colchicine or vinblastine which induces a dose- and time-dependent particle aggregation. These observations suggest that microtubules and other membrane-associated colchicine-sensitive proteins probably influence the distribution of intrinsic membrane proteins and intramembranous particles in nucleated mammalian cells. An aggregated particle distribution has been observed in 3T3 cells or colchicine-treated transformed cells frozen in media, phosphate-buffered saline or following brief exposure to glycerol, sucrose or dimethyl sulfoxide containing solutions, independent of whether specimens were rapidly frozen from 37 degrees C, room temperature or 4 degrees C incubations. Cells briefly stabilized in 1% formaldehyde yields similar patterns of particle distribution as cells rapidly frozen in media or cryoprotectants. Glutaraldehyde fixation of cells, however, appears to alter the fracturing process in these cells, as visualized by an altered fracture face appearance, decreased numbers of particles, and no particle aggregates. Differences in membrane organization between normal and transformed cells have therefore been demonstrated using a series of preparative methods and colchicine and vinblastine have been shown to modulate intramembranous particle distribution in transformed 3T3 cells.
质膜的内在组织在正常细胞和转化细胞中有所不同。通过冷冻断裂和电子显微镜技术,已证明接触抑制的3T3细胞含有聚集的质膜内颗粒,而转化细胞则呈现均匀的颗粒分布。秋水仙碱或长春花碱可影响转化细胞中膜内颗粒的分布,这两种药物会诱导剂量和时间依赖性的颗粒聚集。这些观察结果表明,微管和其他与膜相关的秋水仙碱敏感蛋白可能会影响有核哺乳动物细胞中内在膜蛋白和膜内颗粒的分布。在3T3细胞或在培养基、磷酸盐缓冲盐溶液中冷冻的秋水仙碱处理的转化细胞中,或在短暂暴露于含甘油、蔗糖或二甲基亚砜的溶液后,均观察到聚集的颗粒分布,而与标本是从37℃、室温还是4℃孵育中快速冷冻无关。在1%甲醛中短暂稳定的细胞产生的颗粒分布模式与在培养基或冷冻保护剂中快速冷冻的细胞相似。然而,细胞经戊二醛固定后,似乎会改变这些细胞的断裂过程,这通过断裂面外观改变、颗粒数量减少和无颗粒聚集得以体现。因此,使用一系列制备方法已证明正常细胞和转化细胞之间的膜组织存在差异,并且已证明秋水仙碱和长春花碱可调节转化的3T3细胞中的膜内颗粒分布。