Distribution of fibrillar centers and silver-stained components in the nucleolus of human Sertoli cells

The nucleolus of the human Sertoli cell consistently showed three distinct, spontaneously segregated parts: 1. one or two large, silver-positive fibrillar centers; 2. strands of dense fibrillar component continuous with the dense cords surrounding the fibrillar center. These components were also silver-positive; 3. a granular, silver-negative mass. These observations show that in the human Sertoli cell the number of fibrillar centers is far lower than the diploid number of NORs. They also suggest that the fibrillar center might contain several NORs in this cell type.     

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)     

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.     

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.     

Three-dimensional reconstructions of nucleolus-organizing regions in PHA-stimulated human lymphocytes

Ultrastructure and three-dimensional distribution of nucleolus-organizing regions have been studied on ultrathin serial sections of PHA-stimulated human lymphocytes. During the 48 hr of activation the size of fibrillar centers (FCs) decreased from 0.6-0.9 microns to 0.2-0.3 microns and the number of FCs increased rapidly from one to 75-107 per cell. The number of fibrillar complexes (i.e. associations of a different number of FCs connected by the dense fibrillar component) also increased but did not reach the maximum number of nucleolar organizers presented here. Three-dimensional computer reconstructions of fibrillar complexes showed that lymphocyte activation was accompanied by early (2-4 hr) changes in the shape of the primary fibrillar center. Invagination of the dense fibrillar component on its surface occurred and division into two or more smaller FCs followed. Gradually, the typical structure of the nucleolus with several fibrillar complexes and many FCs was formed. These results confirm the hypothesis of fibrillar complex-nucleolar organizer correlation published recently.     

In situ hybridization at the electron microscope level: an improved method for precise localization of ribosomal DNA and RNA

In situ hybridization using biotinylated rDNA probes and secondary antibody coupled to gold particles was developed on ultrathin sections of Lowicryl-embedded Ehrlich tumor cells for precise localization of ribosomal RNA (rRNA) and ribosomal DNA (rDNA). For the detection of rDNA, an immunocytochemical approach involving an antibody against single-stranded DNA was used in order to determine the more efficient denaturation procedure. Using this technique, rDNA can be visualized in the fibrillar centers of nucleoli, especially in their peripheral regions at the proximity of both the dense fibrils and the nucleolar interstices as well as within the latter. rDNA was occasionally detected in some clumps of dense nucleolus-associated chromatin. Besides the presence of rRNA in the ribosome-rich cytoplasmic areas and in the dense fibrillar component and the granular component of the nucleolus, rRNA was also found in the fibrillar center areas close to the boundary region to the dense fibrillar component. These results are discussed in the light of the present knowledge on the functional organization of the nucleolus.     

New data concerning the functional organization of the mammalian cell nucleolus: detection of RNA and rRNA by in situ molecular immunocytochemistry.

We have investigated the fine spatial distribution of RNA and rRNA within the Ehrlich tumor cell nucleolus by in situ hybridization with a biotin-labeled probe and by two new strategies, the polyadenylate nucleotidyl transferase-immunogold technique and immuno-labeling with anti-RNA antibodies. Besides the presence, as expected, of RNA and rRNA in the granular component and the dense fibrillar component, we show, for the first time, significant label over all the fibrillar centers of the nucleoli. When RNA and DNA were detected simultaneously on the same sections, only the fibrillar centers were positive for both. These results throw light on the controversial subject of the precise location of transcribing rRNA genes within the nucleolus. The fibrillar centers, and not the dense fibrillar component, should thus be the site of rRNA synthesis.     

煤绒菌Fuligo septica显型原质团细胞核及菌核的超微结构

从煤绒菌显型原质团中提纯细胞核、核仁,诱导原质团形成菌核,并在透射电镜下观察。研究结果表明:细胞核具有中央核仁,核仁可以看到明显的纤维中心、致密纤维中心和颗粒结构。原质团中存在大量的黏液颗粒。菌核具有双层膜,内含有细胞器及脂滴。     

Location of DNA within the nucleolus of rat oocytes during the early stages of follicular growth.

We have investigated the DNA distribution within the rat oocyte nucleolus during the early stages of follicular growth by means of the in situ terminal deoxynucleotidyl transferase method. In the fibrillogranular nucleolus, label is visualized on small clumps of peri- and intranucleolar chromatin. Such labeled clumps are frequently observed inside the interstices surrounding the fibrillar centers. Label is also consistently found in the fibrillar centers whereas the dense fibrillar component and the granular component are devoid of gold particles. These results contradict earlier data but conform with other recent immunocytochemical observations, obtained in nucleoli of a variety of somatic cell types, concerning the correlation between structure and function in the nucleolus.     

A comparative study of the main components of the nucleolus in different populations of rat trophoblast cells during differentiation. I. The nucleoli of the trophoblast cells in the labyrinth section of the placenta

The nucleolus undergoes some steps of structural transformation during differentiation of the labyrinth trophoblast cells. Primarily (on day 13 of gestation) the nucleolar components become rather disjoined. The nucleolus is composed of a loose net of strands of granulofibrillar and dense fibrillar components bearing fibrillar centers (FCs). Strands are separated by large lacunae. This rare-occurring type of nucleoli is replaced on the next (14th) day by the nucleolonemal type and later--by the compact nucleolar type. FCs with dense fibrillar component strands become extended into the masses of granulofibrillar component. Such transformations of nucleolar structure seem to be an expression of a fast-proceeding differentiation of the labyrinth trophoblast cells.     

核仁和核仁蛋白

核仁是位于细胞核内的非膜结构。电子显微镜下的核仁从形态上可以分为三层结构包括纤维中心区(FC)、高密度纤维区(DFC)和颗粒区(GC)。核仁内的蛋白有核糖体蛋白和非核糖体蛋白两种。利用蛋白质组学方法已经鉴定了350多种核仁蛋白,其中包括80多种核糖体蛋白。核仁是核糖体合成的场所,核仁中的非核糖体蛋白对核糖体的生物合成起关键调控作用。核仁不仅是细胞内通讯和核糖体：RNA加工的中心,而且在细胞周期、细胞增殖和衰老中起重要调控作用;核仁也是tRNA、mRNA和其它类型小分子RNA加工的场所。因此核仁是一个多功能的细胞生命活动中心。     

煤绒菌Fuligo septica显型原质团细胞核及菌核的超微结构

从煤绒菌显型原质团中提纯细胞核、核仁,诱导原质团形成菌核,并在透射电镜下观察。研究结果表明:细胞核具有中央核仁,核仁可以看到明显的纤维中心、致密纤维中心和颗粒结构。原质团中存在大量的黏液颗粒。菌核具有双层膜,内含有细胞器及脂滴。     

煤绒菌Fuligo septica显型原质团细胞核及菌核的超微结构

从煤绒菌显型原质团中提纯细胞核、核仁,诱导原质团形成菌核,并在透射电镜下观察。研究结果表明:细胞核具有中央核仁,核仁可以看到明显的纤维中心、致密纤维中心和颗粒结构。原质团中存在大量的黏液颗粒。菌核具有双层膜,内含有细胞器及脂滴。     

多头绒泡菌银染核仁周期的电镜研究

以同步化培养的多头绒泡菌（Physarum poldycephalum Schw.)原生质团为材料,应用整体银染技术,电镜下研究了核仁在细胞周期中的超微结构变化。结果变化：核仁成熟时比较大,位于细胞核中央,核仁内可区分出纤维中心、密集纤维成分和颗粒成分等。前期时,核仁向边缘移动,前期末在近核膜处解体,解体的核仁物质主要呈团块状散开。中期时,解体的核仁物质位于细胞核中央染色体区域的周围,染色体上没有特异的银染区域,染色体周边也看不到银染的“鞘”状结构,但在染色体中可见一些散在的银染大颗粒。末期时,核仁物质与染色体一起到达两极,在子细胞核中与正在解集缩的染色质共存一起,以后核仁物质逐渐汇合并与染色质分开。大约在有丝分裂结束120min后,在细胞核中形成一候 中央位置的大核仁,结果提示,低等真核生物的核仁结构和周期变化与高等真核生物的不完全相同。