Non-isotopic detection of nucleolar transcription in pre-implantation mouse embryos

Kobema, Karel; Landa, V.; Kaňka, Jiří; Pliss, Artem; Eltsov, Michail; Staněk, David; Raška, Ivan

doi:10.1051/rnd:19980110

Cited by 3 publications

(1 citation statement)

References 18 publications

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“…By statistical analysis of more than 100 embryos microinjected with BrUTP/␣-amanitin, we could show that nascent pre-rRNAs are first observed at the middle-end of the second cell cycle, around 44 -45 hphCG, and simultaneously in the 2 blastomeres. Transcription sites are associated with the surface of NPBs, in agreement with previous data (Koberna et al, 1998), and we showed here for the first time that they are also associated with small UBF/pol I (RPA116) foci, which decorate the NPBs as necklace-like structures, and thus represent ribosomal gene-bound transcription complexes. Furthermore, the major transcription factors are actually assembled at their rDNA targets at 40 hphCG, i.e., 4 -5 h before the onset of rDNA transcription.…”

Section: Discussionsupporting

confidence: 92%

The Step-Wise Assembly of a Functional Nucleolus in Preimplantation Mouse Embryos Involves the Cajal (Coiled) Body

Зацепина

Baly

Chebrout

et al. 2003

Developmental Biology

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After fertilization, ribosomal RNA synthesis is silenced during a period which depends on the species. Data concerning the reassembly of a functional nucleolus remain scarce. We have examined by immunocytochemistry, Western blots, and BrUTP microinjection the dynamics of major nucleolar proteins during the first cycles of mouse embryogenesis, in relation to rDNA transcription sites and coilin, a marker of Cajal bodies. We show that: (1) the reinitiation of rDNA transcription occurs at the two-cell stage, 44-45 h after hCG injection (hphCG), at the surface of the nucleolar precursor bodies (NPBs), where the RNA polymerase I (pol I) transcription complex is recruited 4-5 h before; (2) the NPBs are not equal in their ability to support recruitment of pol I and rDNA transcription; (3) maternally inherited fibrillarin undergoes a dynamic redistribution during the second cell stage, together with coilin, leading to the assembly of the Cajal body around 40 hphCG; and (4) the pol I complex is first recruited to the Cajal body before reaching its rDNA template. We also find that fibrillarin and B23 are both directly assembled around NPBs prior to ongoing pre-rRNA synthesis. Altogether, our results reveal a role of the Cajal bodies in the building of a functional nucleolus.

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Section: Discussionsupporting

confidence: 92%

The Step-Wise Assembly of a Functional Nucleolus in Preimplantation Mouse Embryos Involves the Cajal (Coiled) Body

Зацепина

Baly

Chebrout

et al. 2003

Developmental Biology

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A non-isotopic assay uses bromouridine and RNA synthesis to detect DNA damage responses

Hasegawa¹,

Iwai²,

Kuraoka³

2010

Mutation Research/Genetic Toxicology and Environmental Mutagene

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Three-Dimensional Distribution of UBF and Nopp140 in Relationship to Ribosomal DNA Transcription During Mouse Preimplantation Development1

Koné

Fleurot

Chebrout

et al. 2016

Biology of Reproduction

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The nucleolus is a dynamic nuclear compartment that is mostly involved in ribosome subunit biogenesis; however, it may also play a role in many other biological processes, such as stress response and the cell cycle. Mainly using electron microscopy, several studies have tried to decipher how active nucleoli are set up during early development in mice. In this study, we analyzed nucleologenesis during mouse early embryonic development using 3D-immunofluorescent detection of UBF and Nopp140, two proteins associated with different nucleolar compartments. UBF is a transcription factor that helps maintain the euchromatic state of ribosomal genes; Nopp140 is a phosphoprotein that has been implicated in pre-rRNA processing. First, using detailed image analyses and the in situ proximity ligation assay technique, we demonstrate that UBF and Nopp140 dynamic redistribution between the two-cell and blastocyst stages (time of implantation) is correlated with morphological and structural modifications that occur in embryonic nucleolar compartments. Our results also support the hypothesis that nucleoli develop at the periphery of nucleolar precursor bodies. Finally, we show that the RNA polymerase I inhibitor CX-5461: 1) disrupts transcriptional activity, 2) alters preimplantation development, and 3) leads to a complete reorganization of UBF and Nopp140 distribution. Altogether, our results underscore that highly dynamic changes are occurring in the nucleoli of embryos and confirm a close link between ribosomal gene transcription and nucleologenesis during the early stages of development.

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Non-isotopic detection of nucleolar transcription in pre-implantation mouse embryos

Abstract: -Nucleolar transcription was analysed in permeabilized pre-implantation mouse embryos at the four-cell, eight-cell, morula and early blastocyst stages using confocal microscopy to detect incorporated 5-

Cited by 3 publications

References 18 publications

The Step-Wise Assembly of a Functional Nucleolus in Preimplantation Mouse Embryos Involves the Cajal (Coiled) Body

The Step-Wise Assembly of a Functional Nucleolus in Preimplantation Mouse Embryos Involves the Cajal (Coiled) Body

A non-isotopic assay uses bromouridine and RNA synthesis to detect DNA damage responses

Three-Dimensional Distribution of UBF and Nopp140 in Relationship to Ribosomal DNA Transcription During Mouse Preimplantation Development1

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