Ferreira J, Carmo-Fonseca M
Institute of Histology and Embryology, Faculty of Medicine, University of Lisbon, Portugal.
Development. 1995 Feb;121(2):601-12. doi: 10.1242/dev.121.2.601.
The coiled body is an ubiquitous nuclear organelle that contains essential components of the pre-mRNA splicing machinery as well as the nucleolar protein fibrillarin. Here we have studied the biogenesis of the coiled body in early mouse embryos. The results show that coiled bodies form and concentrate splicing snRNPs as early as in the maternal and paternal pronuclei of 1-cell embryos. This argues that the coiled body is likely to play a basic role in the nucleus of mammalian cells. In order to correlate the appearance of coiled bodies with the onset of transcriptional activity, embryos were incubated with brominated UTP and the incorporated nucleotide was visualized by fluorescence microscopy. In agreement with previous studies, transcriptional activity was first observed during the 2-cell stage. Thus, coiled bodies form before activation of embryonic gene expression. The appearance of coiled bodies in 1-cell embryos was preceded by the formation of morphologically distinct structures that also contain coilin and which we therefore refer to as pre-coiled bodies. At the electron microscopic level pre-coiled bodies have a compact fibrillar structure, whereas coiled bodies resemble a tangle of coiled threads. Although both pre-coiled bodies and coiled bodies contain the nucleolar protein fibrillarin, the assembly of coiled bodies is separated both in time and in space from ribosome synthesis. Our results suggest that the embryonic 'nucleolus-like body' is a structural scaffold that nucleates independently the formation of the coiled body and the assembly of the machinery responsible for ribosome biosynthesis.
卷曲小体是一种普遍存在的核细胞器,它包含前体mRNA剪接机制的重要成分以及核仁蛋白纤维蛋白。在此,我们研究了小鼠早期胚胎中卷曲小体的生物发生过程。结果表明,早在1细胞胚胎的雄原核和雌原核中,卷曲小体就开始形成并浓缩剪接snRNP。这表明卷曲小体可能在哺乳动物细胞核中发挥基本作用。为了将卷曲小体的出现与转录活性的开始联系起来,将胚胎与溴化UTP一起孵育,并通过荧光显微镜观察掺入的核苷酸。与先前的研究一致,转录活性最早在2细胞阶段观察到。因此,卷曲小体在胚胎基因表达激活之前就已形成。在1细胞胚胎中卷曲小体出现之前,先形成了形态上不同的结构,这些结构也含有卷曲蛋白,因此我们将其称为前卷曲小体。在电子显微镜水平上,前卷曲小体具有紧密的纤维状结构,而卷曲小体则类似于一团盘绕的线。尽管前卷曲小体和卷曲小体都含有核仁蛋白纤维蛋白,但卷曲小体的组装在时间和空间上都与核糖体合成分开。我们的结果表明,胚胎“核仁样体”是一种结构支架,它独立地促使卷曲小体的形成以及负责核糖体生物合成的机制的组装。