Quantitative ultrastructural study of nucleolus-organizing regions at some stages of the cell cycle (G0 period, G2 period, mitosis)

The quantitative characteristics of chromosomal nucleolus-organizing regions (NORs) and some other nucleolar components were studied on ultra-thin sections of pig embryo kidney cells (PK cells). It was shown that: 1) nucleoli-per-cell volumes were 3 times smaller in the G0 period than in the G2 period; 2) the number of fibrillar centers (FCs) per cell in the G0 period, the G2 period, and at metaphase was equal to 7, 33.7, and 8, respectively; 3) mean volumes of individual FCs in the G0 period (0.033 +/- 0.005 micron3), G2 period (0.014 +/- 0.001 micron3), and at metaphase (0.025 +/- 0.002 micron3) were significantly different; 4) the total volumes of FCs calculated per haploid set of chromosomes were practically the same in the G0 (0.105 micron3) and G2 (0.107 micron3) periods, but were twice as large as those at metaphase (0.04-0.05 micron3). These data show that partial activation and inactivation of ribosomal genes in interphase PK cells are not accompanied by a considerable change in the total volume of FCs and may be due to the fragmentation and fusion of individual FCs. Complete inactivation of ribosomal genes in mitosis results in a decrease of total volumes of FCs per cell; 5) in G0 and G2 periods the total volume of the dense fibrillar component per nucleolus is practically proportional to the nucleolus volume (r = 0.99); 6) in the G2 period, the nucleolus volume is also proportional to the number of FCs (r = 0.99; 7) the volume of the dense fibrillar component within individual fibrillar complexes is not a constant one.(ABSTRACT TRUNCATED AT 250 WORDS)     

Morphometric study of nucleolus-organizing regions of chromosomes from swine embryo kidney cells during Go-, G2-period and mitosis

Quantitative characteristics of nucleolus-organizing regions of chromosomes (NORs, or fibrillar centers, FCs) and some other nucleolar components have been studied with the aid of complete series of ultrathin sections of PK-cells. It has been found that: 1) the number of FCs per cell in the G0-period, in the G2-period and at metaphase is equal to 7.0, 33.7 and 8.0, respectively; 2) volumes of individual FCs in the G0-period (0.033 micron 3), G2-period (0.014 micron 3) and at metaphase (0.025 micron 3) are different; 3) the total volume of FCs, calculated for a haploid set of chromosomes, do not differ in the G0 (0.105 micron 3) and G2 (0.107 micron 3) periods, but exceed twice the FCs volume at metaphase (0.04-0.05 micron 3). These data show that the activation and inactivation of ribosomal genes in interphase PK-cells are not accompanied by a change in the total volumes of FCs and are probably connected with the "fragmentation" and fusion of FCs. Complete inactivation of ribosomal genes at mitosis leads to a decrease in the total volumes of FCs; 4) the nucleolus volume is proportional to the volume of the dense fibrillar RNP-component; in the G2-period the nucleolus volume also correlates with the number of FCs (r = 0.99); 5) the volume of the dense fibrillar component within individual fibrillar complexes--the structures corresponding to one nucleolus-organizing region--is not constant. This is an indirect evidence for the differences in the functional activity of NORs of different chromosomes.     

Subnucleolar distribution and organization of Vicia faba L. rDNA in situ

The distribution and organization of nucleolar DNA in Vicia faba L. was analyzed by specific cytochemical staining using NAMA-Ur. The results showed that nucleolar DNA was distributed in the FCs and at the FC/DFC junctions. Statistical analysis showed that the rRNA genes occupied about one-third of the total dense fibrillar component region. The rDNA was condensed in some regions and uncondensed in others. Nucleolus-associated chromatin extended from outside the nucleolus to the periphery of the FCs via nucleolar channels, suggesting a possible origin for nucleolar DNA.     

Localization of U3 RNA molecules in nucleoli of HeLa and mouse 3T3 cells by high resolution in situ hybridization.

We have examined the ultrastructural localization of U3 RNA in the nucleoli of HeLa and mouse 3T3 cells by in situ hybridization with a biotinylated U3 DNA probe and subsequent detection of hybrids with electron microscopy by direct immunogold labeling. The highest levels of signal density for U3 RNA are detected over the dense fibrillar component (DFC) of the nucleolus, including the interfaces between DFC and the enclosed fibrillar center (FC) on the one hand and DFC and the granular component (GC) on the other hand. Lower but significant signals also are observed over GC, which indicate, taking into account the high relative volume of GC in a nucleolus, that a substantial fraction of U3 RNA is present in this compartment where the more mature forms of pre-rRNA accumulate. In parallel, the localization of fibrillarin was analyzed by immunogold detection, demonstrating that fibrillarin and U3 RNA have a roughly similar distribution, although quantitative measurements reveal that the signal ratio for both molecules exhibit significant differences among the major ultrastructural components of the nucleolus.     

Studies on changes in nucleolar organizer region of human promyelocytic leukemia cells (HL-60) treated with retinoic acid

Changes of nucleolar organizer region in HL-60 cells after treated with retinoic acid (RA) were studied with techniques of silver-staining nucleolar organizer region (Ag-NOR) in metaphase karyotypes, Brachet's reaction and with our improved TEM techniques for studying silver-stained active nucleolar organizer region (Ag-aNOR) in interphase nucleoli. Number of Ag-NOR in HL-60 cells is 4.5/cell on average. The Ag-NOR number of cells treated with RA showed no remarkable difference from that of control group. Ag-aNOR number treated with RA was reduced obviously as compared with that of control group. Meanwhile, the changes of nucleolus number showed by Brachet's reaction were in accordance with those of Ag-aNOR. Therefore, it may be concluded: (1). Though the number of active rRNA genes did not changed after the differentiation of HL-60 cells induced by RA, their expression was clearly inhibited: (2). The relationship between the changes of Brachet-No and Ag-aNOR is in positive correlation (r = 0.98, p less than 0.01). EM examination of Ag-aNOR of HL-60 cells reveals that Ag-protein (RNA polymerase I) only presented in fibrillar centers (FC) and the dense fibrillar components (DFC) of nucleolus. In addition, in control group, large amount of Ag-protein, FC, DFC and granular components (GC) were observed, and there were many large nucleoli in a nucleus, meanwhile, the cells of the treated group tended to be mature, with a decrease in the amount of Ag-protein, FC, DFC and GC accordingly, and the nucleoli reduced both in size and number significantly.(ABSTRACT TRUNCATED AT 250 WORDS)     

洋葱细胞核仁内DNA的结构变化及排布过程(英文)

以洋葱 (AlliumcepaL .)细胞为研究材料 ,应用DNA细胞化学特异染色方法 (NAMA_Ur)及常规电子显微镜技术 ,观察了洋葱细胞核仁FC(纤维中心 )内DNA的超微结构 ,发现FC内DNA存在着一个介于集缩到解集缩之间的变化过程 ,并揭示了DNA在核仁内的连续排布过程 ,即核仁外DNA经过核仁通道进入到FC后 ,继续沿FC的边缘或DFC(致密纤维成分 )与FC的交界处环绕FC而排布 ,再经FC之间的核仁通道 ,延伸到另外的FC区域     

The Structural Transformation of Nucleolar DNA and Its Arrangement in Nucleolus of Allium cepa Cells

By using the DNA specific cytochemical staining method (NAMA-Ur) and conventional electron microscopic technique, the authors examined the configuration of intranucleolar DNA in Allium cepa L. cells and found that nucleolar DNA within the fibrillar center (FC) underwent a structural transformation process from condensed to extended state. The authors' observations also displayed a continuous arrangement process of nucleolar DNA, i.e., the extranucleolar DNA entered FC through the nucleolar organizer region (NOR) channel, then extended to the periphery of FC or to the border between FC and dense fibrillar component (DFC), and distributed along the periphery of FC. Thence, by passing through the NOR channel between FCs, the nucleolar DNA continued to transfer to other FCs and arranged in the same above-mentioned forms.