Manuelidis L
J Neuropathol Exp Neurol. 1984 May;43(3):225-41. doi: 10.1097/00005072-198405000-00002.
Active nucleolus organizing regions (NOR) were stained with silver in isolated nuclei and chromosomes, monolayer cultures, and tissues for electron microscopy. In nuclei isolated in 0.8-1.3 M urea, only the NOR stained with silver, and round or fiber-like regions with a diameter of 200-250 nm with a substructure were delineated at appropriate nucleolar or acrocentric chromosomal sites. In whole cells, additional (non NOR) silver binding regions were noted. Electrophoresis studies on nuclear isolates indicated at least two different nuclear protein subsets were responsible for the observed silver binding; the NOR protein(s) was most tightly bound with respect to urea. Interphase cells displayed a more extensive NOR network than mitotic cells, suggesting an increase in rDNA transcription during interphase. Mature sperm cells showed no active NOR regions. Interphase neuroectodermal tumor cells generally contained large multiple NOR indicating extensive activity; the position of NOR in the nucleus was variable from cell to cell, even in cloned lines. In contrast to tumor lines, central nervous system (CNS) cells from various species all showed highly reproducible or non-random NOR locations within the nucleus of each cell type. The NOR of large neurons were always central and single, whereas small granule cell neurons displayed a few small NOR that were positioned more peripherally. These findings suggest that in highly differentiated cells, the NOR region is precisely positioned in the nucleus, regardless of non-identical chromosome locations of the NOR in different species. The variability of NOR in tumor cells indicates profound changes in nuclear structure that may be part of the neoplastic transformation.
在分离的细胞核、染色体、单层培养细胞和用于电子显微镜观察的组织中,活性核仁组织区(NOR)用银进行染色。在0.8 - 1.3M尿素中分离的细胞核中,只有NOR被银染色,并且在适当的核仁或近端着丝粒染色体位点勾勒出直径为200 - 250nm且具有亚结构的圆形或纤维状区域。在全细胞中,还观察到其他(非NOR)银结合区域。对核分离物的电泳研究表明,至少有两种不同的核蛋白亚群负责观察到的银结合;NOR蛋白相对于尿素结合最紧密。间期细胞显示出比有丝分裂细胞更广泛的NOR网络,表明间期rDNA转录增加。成熟精子细胞未显示出活性NOR区域。间期神经外胚层肿瘤细胞通常含有大量多个NOR,表明活性广泛;即使在克隆系中,NOR在细胞核中的位置在细胞之间也是可变的。与肿瘤细胞系不同,来自各种物种的中枢神经系统(CNS)细胞在每种细胞类型的细胞核内都显示出高度可重复或非随机的NOR位置。大神经元的NOR总是位于中央且单一,而小颗粒细胞神经元显示出一些位于更外周的小NOR。这些发现表明,在高度分化的细胞中,无论NOR在不同物种中的染色体位置是否相同,NOR区域在细胞核中都精确定位。肿瘤细胞中NOR的变异性表明核结构发生了深刻变化,这可能是肿瘤转化的一部分。