Moreno F J, Rodrigo R M, Garcia-Herdugo G
Department of Cell Biology, Faculty of Biology, University of Seville, Spain.
J Histochem Cytochem. 1990 Dec;38(12):1879-87. doi: 10.1177/38.12.1701461.
In this work we used Allium cepa root meristem cells in actively growing conditions and under treatment with the protein synthesis inhibitors cycloheximide (CHM) and puromycin. Morphological and quantitative results indicate that these drugs induce dramatic alterations in nucleolar structure reflected by a decrease of nucleolar size, much more evident under treatment with CHM, and by segregation of its main components. Quantitative analysis shows a decrease in NOR-silver staining after treatment with CHM, whereas in cells treated with puromycin NOR-silver staining remains constant. Our results reveal a decrease in the Ag-NOR proteins under conditions of diminished cell activity, suggesting a direct relationship between the quantity of Ag-NOR proteins and transcriptional activity. Using two-dimensional gel electrophoresis and NOR-silver staining in gels, we have characterized some proteins corresponding to molecular weights of 28 and 31 KD and pI of approximately 5.2. After treatment with CHM, reactivity of these proteins against NOR-silver staining is diminished. By means of a morphological study, analysis of NOR-silver staining, and of anti-DNA and RNAse-gold labeling, we have tried an approach to the nucleolar organization in plant cells. Our results suggest that the fibrillar component shows a reticular distribution where fibrillar centers, as described in animal cells, are not distinguished.
在这项工作中,我们使用了处于活跃生长状态且用蛋白质合成抑制剂环己酰亚胺(CHM)和嘌呤霉素处理的洋葱根尖分生组织细胞。形态学和定量结果表明,这些药物会诱导核仁结构发生显著变化,表现为核仁大小减小,在CHM处理下更为明显,以及其主要成分的分离。定量分析显示,用CHM处理后NOR银染减少,而用嘌呤霉素处理的细胞中NOR银染保持不变。我们的结果揭示了在细胞活性降低的情况下Ag-NOR蛋白减少,这表明Ag-NOR蛋白的数量与转录活性之间存在直接关系。通过二维凝胶电泳和凝胶中的NOR银染,我们鉴定了一些分子量为28和31 KD、pI约为5.2的蛋白质。用CHM处理后,这些蛋白质对NOR银染的反应性降低。通过形态学研究、NOR银染分析以及抗DNA和RNA酶-金标记,我们尝试了一种研究植物细胞核仁组织的方法。我们的结果表明,纤维成分呈网状分布,其中未区分出动物细胞中所述的纤维中心。