The role of RNA polymerase I transcription and embryonic genome activation in nucleolar development in bovine preimplantation embryos

Molecular Reproduction and Development
Olga SvarcovaPoul Maddox-Hyttel

Abstract

The aim of the present study was to investigate the role of RNA polymerase I (RPI) transcription in nucleolar development during major transcriptional activation (MTA) in cattle. Late eight-cell embryos were cultured in the absence (control group) or presence of actinomycin D (AD) (RPI inhibition, AD 0.2 microg/ml; total transcriptional inhibition, AD 2.0 microg/ml). Late four-cell embryos were cultured to late eight-cell stage in 0.2 microg/ml AD (MTA prevention, ADLT (long-term total transcriptional inhibition group). Embryos were processed for autoradiography, transmission electron microscopy, fluorescent in situ hybridization (ribosomal RNA, rRNA), silver staining (nucleolar proteins), and immunofluorescence (RPI). Control embryos displayed extranucleolar and nucleolar transcription, functional nucleoli, and distinct RPI localization. Nuclei (97%) showed large rRNA clusters, in 94.1% co-localized with nucleolar proteins deposits. In AD 0.2 group, only extranucleolar transcription was detected. Segregated dense-fibrillar and granular components, but no fibrillar centers, were observed. RPI was dispersed. Nuclei (55%) presented rRNA clusters, in 38.8% co-localized with silver-stained deposits. AD 2.0 and ADLT groups displayed...Continue Reading

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Citations

Mar 9, 2012·Planta·Katarzyna NiedojadłoElżbieta Bednarska-Kozakiewicz
Jul 15, 2009·Cloning and Stem Cells·Olga ØstrupPoul Maddox-Hyttel
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May 13, 2008·Molecular Reproduction and Development·O SvarcovaP Maddox-Hyttel

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