Kopecny V, Biggiogera M, Pivko J, Grafenau P, Pavlok A, Malatesta M, Martin T E, Fakan S
Centre of Electron Microscopy, University of Lausanne, Switzerland.
Eur J Cell Biol. 1996 Aug;70(4):361-72.
Fine structural cytochemistry and immunocytochemistry were used to study nucleic acids and nuclear proteins in nuclear bodies (NB) of pronuclear and 2-cell bovine and caprine embryos on ultrathin sections of paraformaldehyde fixed and Lowicryl K4M or LR White embedded specimens. The most striking feature detected in some of these nuclear bodies (NBs) was the presence of non-nucleolar proteins known to be involved in pre-mRNA splicing. One category of such intranuclear bodies (showing a rather dense finely fibrillar composition and named here dense body-DB) contained the Sm-antigen (an antigen common to a major group of nucleoplasmic spliceosomal snRNPs). Another, more numerous category of NBs differed morphologically from the former one by a much looser composition of fibrillogranular elements (loose body-LB). Moreover, it showed the presence of the non-snRNP splicing factor SC-35, in addition to the Sm-antigen. Both categories of these nuclear bodies were distinguished clearly from the nucleolar precursor bodies (NPBs) by an absence of immunolabeling of NPB with antibodies against nuclear proteins involved in splicing. Moreover, the former NBs are not stained with silver, while NPBs already in pronuclei exhibit strong affinity to silver. In addition to the immunolabeling in prominent (approx. 0.2-2.0 microns) NBs, regularly occurring high concentration of snRNP was revealed in very small (approx. 0.05 micron), morphologically poorly defined areas (named here small snRNP-enriched areas-SSA), harboring moreover a set of nuclear proteins similar to that of the coiled body. Numerous observations of the presence of these small areas in nuclear bodies and in their close vicinity, in nucleoplasm, in proximity of the nuclear envelope and also in ooplasm suggested that they are possible carriers of certain nuclear proteins moving between nuclear bodies, nucleoplasm and cytoplasm. A functional relationship of all these embryonic subnuclear elements has not been elucidated so far but their mutual relation is suggested, since the NPBs and other nuclear bodies usually occur in a close association. Fine structural and immunoelectron microscopic observations further suggest a similarity of the nuclear bodies in the early ruminant embryo with specific intranuclear bodies ("snurposomes") known from Xenopus laevis oocytes. A new and striking feature emerging from these observations is a possible involvement of a group of nucleoplasmic proteins in a yet unknown way in the differentiation processes concomitant with early embryonic nucleologenesis.
利用精细结构细胞化学和免疫细胞化学技术,对经多聚甲醛固定、用Lowicryl K4M或LR White包埋的原核期和2细胞期牛和羊胚胎超薄切片中的核小体(NB)内的核酸和核蛋白进行研究。在其中一些核小体(NBs)中检测到的最显著特征是存在已知参与前体mRNA剪接的非核仁蛋白。一类这样的核内体(显示出相当致密的细纤维成分,在此命名为致密体-DB)含有Sm抗原(一种主要的核质剪接体snRNP共有的抗原)。另一类数量更多的核小体在形态上与前者不同,其纤维颗粒成分更为疏松(疏松体-LB)。此外,除了Sm抗原外,它还显示存在非snRNP剪接因子SC-35。这两类核小体通过用针对参与剪接的核蛋白的抗体对核仁前体小体(NPB)进行免疫标记时未出现免疫反应,与核仁前体小体(NPB)明显区分开来。此外,前者的核小体不被银染色,而原核中的核仁前体小体对银具有很强的亲和力。除了在明显的(约0.2 - 2.0微米)核小体中有免疫标记外,在非常小的(约0.05微米)、形态上界定不清的区域(在此命名为富含snRNP的小区域-SSA)中还发现了经常出现的高浓度snRNP,此外该区域还含有一组与卷曲体相似的核蛋白。在核小体及其附近、核质中、靠近核膜处以及卵质中多次观察到这些小区域的存在,这表明它们可能是某些在核小体、核质和细胞质之间移动的核蛋白的载体。目前尚未阐明所有这些胚胎亚核元件之间的功能关系,但由于核仁前体小体(NPBs)和其他核小体通常紧密相连,因此提示了它们之间的相互关系。精细结构和免疫电子显微镜观察进一步表明,反刍动物早期胚胎中的核小体与非洲爪蟾卵母细胞中已知的特定核内体(“snurposomes”)相似。这些观察结果中出现的一个新的显著特征是,一组核质蛋白可能以一种未知的方式参与了与早期胚胎核仁形成相关的分化过程。