Anisimova A A, Anisimov A P
Tsitologiia. 2005;47(1):14-22.
Variation of some characteristics of nucleoli of polyploid mucous and albumen cells was examined in salivary glands of the snail Succinea lauta. The number, total area and Ag-protein content of nucleoli, and DNA content in each nucleus were estimated on squashed preparations incubated with AgNO3, decolorized and then Feulgen stained. The ultrastructure of nucleoli was studied by electron microscopy. Differentiated mucous cells had 4c-8c-16c-32c nuclei; albumen cells had 8c-16c-32c-64c-128c nuclei. The ultrastructure of nucleoli of the two cell types was essentially the same. Normally, a large fibrous to granular zone was observed in the nucleoli, without a clear distinction between fibrous and granular components. At the same time, aggregations of granular matter could be discerned at the periphery of nucleoli. No fibrous centers were observed. Occassionally, nucleolonema-like structures occurred. Normally each nucleolus contacted several chromosomes. On squashed preparations, the least size of nucleoli was 2-3 microm, and the largest size amounted to 14 microm in mucous cells, and to 50-80 microm in albumen cells. The number of nucleoli rose from 1-2 in tetraploid nuclei to 2-3 in 32c-nuclei, and to 5-7 in 128c-nuclei. The disparity between the ploidy levels of nuclei and the numbers of nucleoli may be due, presumably, to aggregation of chromosome NORs. The Ag-protein content in the nucleoli, and the total nucleolar area displayed a strong mutual correlation. Both parameters differed significantly by 1.5-2.2 times in mucous and albumen cells of the same ploidy level. Thus, in albumen and mucous cells the total Ag-protein content in octaploid nuclei was 3.3 and 2.2 relative units (r. u.), respectively. In 16c- and 32c-nuclei of albumen cells, it was 7.6 and 15.1 r. u.; and in the same nuclei of mucous cells--3.8 and 6.8 r. u., respectively. On the whole, in albumen cells, in the course of 4 endocycles (4c-128c), the total Ag-protein content increased by 17 times. Therefore, the mean multiplication factor for this parameter was found to be 2.05 per endocycle. In mucous cells, in the course of 3 endocycles (4c-32c), the total Ag-protein content increased by 5.2 times against 8 times expected, with the mean multiplication factor equal to 1.75 per endocycle. Thus, in the course of polyploidization of albumen and mucous cell nuclei, the gene dosage effect was fully pronounced in the former, and only partly in the latter. This differtence is due obviously to peculiarities of differentiation of the two cell types, in particular, to differences in the number of activated ribosomal genes.
研究了苏氏琥珀螺唾液腺中多倍体黏液细胞和蛋白细胞的核仁某些特征的变化。在经硝酸银孵育、脱色然后进行福尔根染色的压片标本上,估计了核仁的数量、总面积和银蛋白含量以及每个细胞核中的DNA含量。通过电子显微镜研究了核仁的超微结构。分化的黏液细胞具有4c - 8c - 16c - 32c的细胞核;蛋白细胞具有8c - 16c - 32c - 64c - 128c的细胞核。两种细胞类型核仁的超微结构基本相同。正常情况下,在核仁中观察到一个大的纤维状到颗粒状区域,纤维状和颗粒状成分之间没有明显区别。同时,在核仁周边可以辨别出颗粒物质的聚集。未观察到纤维中心。偶尔会出现类核仁丝结构。正常情况下每个核仁与几条染色体接触。在压片标本上,黏液细胞核仁最小尺寸为2 - 3微米,最大尺寸达14微米,而蛋白细胞核仁最大尺寸为50 - 80微米。核仁数量从四倍体细胞核中的1 - 2个增加到32c细胞核中的2 - 3个,再增加到128c细胞核中的5 - 7个。细胞核倍性水平与核仁数量之间的差异可能大概是由于染色体核仁组织区的聚集。核仁中的银蛋白含量与核仁总面积显示出很强的相互相关性。在相同倍性水平的黏液细胞和蛋白细胞中,这两个参数有显著差异,相差1.5 - 2.2倍。因此,在八倍体细胞核中,蛋白细胞和黏液细胞的总银蛋白含量分别为3.3和2.2相对单位(r.u.)。在蛋白细胞的16c和32c细胞核中,分别为7.6和15.1 r.u.;而在黏液细胞的相同细胞核中,分别为3.8和6.8 r.u.。总体而言,在蛋白细胞中,在4次内循环(4c - 128c)过程中,总银蛋白含量增加了17倍。因此,该参数的平均倍增系数为每个内循环2.05。在黏液细胞中,在3次内循环(4c - 32c)过程中,总银蛋白含量增加了5.2倍,而预期增加8倍,平均倍增系数为每个内循环1.75。因此,在蛋白细胞和黏液细胞核多倍体化过程中,基因剂量效应在前者中充分体现,而在后者中仅部分体现。这种差异显然是由于两种细胞类型分化的特殊性,特别是活化核糖体基因数量的差异。