The biogenesis of the coiled body during early mouse development

Development ◽  
1995 ◽  
Vol 121 (2) ◽  
pp. 601-612 ◽  
Author(s):  
J. Ferreira ◽  
M. Carmo-Fonseca

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.

1994 ◽  
Vol 126 (1) ◽  
pp. 11-23 ◽  
Author(s):  
J A Ferreira ◽  
M Carmo-Fonseca ◽  
A I Lamond

In the interphase nucleus of mammalian cells the U1, U2, U4/U6, and U5 small nuclear ribonucleoproteins (snRNPs), which are subunits of spliceosomes, associate with specific subnuclear domains including interchromatin granules and coiled bodies. Here, we analyze the association of splicing snRNPs with these structures during mitosis and reassembly of daughter nuclei. At the onset of mitosis snRNPs are predominantly diffuse in the cytoplasm, although a subset remain associated with remnants of coiled bodies and clusters of mitotic interchromatin granules, respectively. The number and size of mitotic coiled bodies remain approximately unchanged from metaphase to early telophase while snRNP-containing clusters of mitotic interchromatin granules increase in size and number as cells progress from anaphase to telophase. During telophase snRNPs are transported into daughter nuclei while the clusters of mitotic interchromatin granules remain in the cytoplasm. The timing of nuclear import of splicing snRNPs closely correlates with the onset of transcriptional activity in daughter nuclei. When transcription restarts in telophase cells snRNPs have a diffuse nucleoplasmic distribution. As cells progress to G1 snRNP-containing clusters of interchromatin granules reappear in the nucleus. Coiled bodies appear later in G1, although the coiled body antigen, p80 coilin, enters early into telophase nuclei. After inhibition of transcription we still observe nuclear import of snRNPs and the subsequent appearance of snRNP-containing clusters of interchromatin granules, but not coiled body formation. These data demonstrate that snRNP associations with coiled bodies and interchromatin granules are differentially regulated during the cell division cycle and suggest that these structures play distinct roles connected with snRNP structure, transport, and/or function.


1993 ◽  
Vol 120 (4) ◽  
pp. 841-852 ◽  
Author(s):  
M Carmo-Fonseca ◽  
J Ferreira ◽  
A I Lamond

Coiled bodies (CBs) are nuclear organelles in which splicing snRNPs concentrate. While CBs are sometimes observed in association with the nucleolar periphery, they are shown not to contain 5S or 28S rRNA or the U3 snoRNA. This argues against CBs playing a role in rRNA maturation or transport as previously suggested. We present evidence here that CBs are kinetic structures and demonstrate that the formation of snRNP-containing CBs is regulated in interphase and mitosis. The coiled body antigen, p80 coilin, was present in all cell types studied, even when CBs were not prominent. Striking changes in the formation of CBs could be induced by changes in cellular growth temperature without a concomitant change in the intracellular p80 coilin level. During mitosis, CBs disassemble, coinciding with a mitotic-specific phosphorylation of p80 coilin. Coilin is shown to be a phosphoprotein that is phosphorylated on at least two additional sites during mitosis. CBs reform in daughter nuclei after a lag period during which they are not detected. CBs are thus, dynamic nuclear organelles and we propose that cycling interactions of splicing snRNPs with CBs may be important for their participation in the processing or transport of pre-mRNA in mammalian cells.


Author(s):  
K. Brasch ◽  
J. Williams ◽  
D. Gallo ◽  
T. Lee ◽  
R. L. Ochs

Though first described in 1903 by Ramon-y-Cajal as silver-staining “accessory bodies” to nucleoli, nuclear bodies were subsequently rediscovered by electron microscopy about 30 years ago. Nuclear bodies are ubiquitous, but seem most abundant in hyperactive and malignant cells. The best studied type of nuclear body is the coiled body (CB), so termed due to characteristic morphology and content of a unique protein, p80-coilin (Fig.1). While no specific functions have as yet been assigned to CBs, they contain spliceosome snRNAs and proteins, and also the nucleolar protein fibrillarin. In addition, there is mounting evidence that CBs arise from or are generated near the nucleolus and then migrate into the nucleoplasm. This suggests that as yet undefined links may exist, between nucleolar pre-rRNA processing events and the spliceosome-associated Sm proteins in CBs.We are examining CB and nucleolar changes in three diverse model systems: (1) estrogen stimulated chick liver, (2) normal and neoplastic cells, and (3) polyploid mouse liver.


2008 ◽  
Vol 103 (6) ◽  
pp. 1856-1865 ◽  
Author(s):  
Ipsita Dey-Guha ◽  
Nasir Malik ◽  
Renaud Lesourne ◽  
Paul E. Love ◽  
Heiner Westphal

2009 ◽  
Vol 126 ◽  
pp. S101-S102
Author(s):  
Stefan Rudloff ◽  
Rolf Kemler

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