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

Olga Østrup; F. Strejcek; I. Petrovicova; Birthe Margit Avery; Hanne Gervi Pedersen; A. Lucas-Hahn; H. Niemann; J. Laurincik; Poul Maddox-Hyttel

doi:10.1002/mrd.20865

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

Olga Østrup, F. Strejcek, I. Petrovicova, Birthe Margit Avery, Hanne Gervi Pedersen, A. Lucas-Hahn, H. Niemann, J. Laurincik, Poul Maddox-Hyttel

14 Citations (Scopus)

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 µg/ml; total transcriptional inhibition, AD 2.0 µg/ml). Late four-cell embryos were cultured to late eight-cell stage in 0.2 µg/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. RPPI was dispersed. Nuclei (55%) presented rRNA clusters, in 38.8% co-localized with silver-stained deposits. AD 2.0 and ADLT groups displayed no transcription and disintegrating nucleolar precursors. AD 2.0 (34%) and 14% (ADLT) of nuclei presented clusters of maternally inherited rRNA. In AD 2.0 group, RPI was dispersed, but 17.2% of nuclei showed colocalization of rRNA with nucleolar proteins. In ADLT group, RPI was lacking and clustering of nucleolar proteins was hampered. In conclusion, rDNA transcription is not required for targeting of rRNA processing proteins, rRNA is maternally inherited and target to rDNA independent of transcription, and de novo transcription is required for proper nucleologenesis in cattle.

Original language	English
Journal	Molecular Reproduction and Development
Volume	75
Issue number	7
Pages (from-to)	1095-1103
Number of pages	9
ISSN	1040-452X
DOIs	https://doi.org/10.1002/mrd.20865
Publication status	Published - 2008

Keywords

Former LIFE faculty
RNA polymerase I
bovine embryo
genome activation
nucleolus

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10.1002/mrd.20865

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Østrup, O., Strejcek, F., Petrovicova, I., Avery, B. M., Pedersen, H. G., Lucas-Hahn, A., Niemann, H., Laurincik, J., & Maddox-Hyttel, P. (2008). The role of RNA polymerase I transcription and embryonic genome activation in nucleolar development in bovine preimplantation embryos. Molecular Reproduction and Development, 75(7), 1095-1103. https://doi.org/10.1002/mrd.20865

Østrup, O, Strejcek, F, Petrovicova, I, Avery, BM, Pedersen, HG, Lucas-Hahn, A, Niemann, H, Laurincik, J & Maddox-Hyttel, P 2008, 'The role of RNA polymerase I transcription and embryonic genome activation in nucleolar development in bovine preimplantation embryos', Molecular Reproduction and Development, vol. 75, no. 7, pp. 1095-1103. https://doi.org/10.1002/mrd.20865

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

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 µg/ml; total transcriptional inhibition, AD 2.0 µg/ml). Late four-cell embryos were cultured to late eight-cell stage in 0.2 µg/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. RPPI was dispersed. Nuclei (55%) presented rRNA clusters, in 38.8% co-localized with silver-stained deposits. AD 2.0 and ADLT groups displayed no transcription and disintegrating nucleolar precursors. AD 2.0 (34%) and 14% (ADLT) of nuclei presented clusters of maternally inherited rRNA. In AD 2.0 group, RPI was dispersed, but 17.2% of nuclei showed colocalization of rRNA with nucleolar proteins. In ADLT group, RPI was lacking and clustering of nucleolar proteins was hampered. In conclusion, rDNA transcription is not required for targeting of rRNA processing proteins, rRNA is maternally inherited and target to rDNA independent of transcription, and de novo transcription is required for proper nucleologenesis in cattle.",

keywords = "Former LIFE faculty, RNA polymerase I, bovine embryo, genome activation, nucleolus",

author = "Olga {\O}strup and F. Strejcek and I. Petrovicova and Avery, {Birthe Margit} and Pedersen, {Hanne Gervi} and A. Lucas-Hahn and H. Niemann and J. Laurincik and Poul Maddox-Hyttel",

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doi = "10.1002/mrd.20865",

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TY - JOUR

T1 - The role of RNA polymerase I transcription and embryonic genome activation in nucleolar development in bovine preimplantation embryos

AU - Østrup, Olga

AU - Strejcek, F.

AU - Petrovicova, I.

AU - Avery, Birthe Margit

AU - Pedersen, Hanne Gervi

AU - Lucas-Hahn, A.

AU - Niemann, H.

AU - Laurincik, J.

AU - Maddox-Hyttel, Poul

PY - 2008

Y1 - 2008

N2 - 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 µg/ml; total transcriptional inhibition, AD 2.0 µg/ml). Late four-cell embryos were cultured to late eight-cell stage in 0.2 µg/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. RPPI was dispersed. Nuclei (55%) presented rRNA clusters, in 38.8% co-localized with silver-stained deposits. AD 2.0 and ADLT groups displayed no transcription and disintegrating nucleolar precursors. AD 2.0 (34%) and 14% (ADLT) of nuclei presented clusters of maternally inherited rRNA. In AD 2.0 group, RPI was dispersed, but 17.2% of nuclei showed colocalization of rRNA with nucleolar proteins. In ADLT group, RPI was lacking and clustering of nucleolar proteins was hampered. In conclusion, rDNA transcription is not required for targeting of rRNA processing proteins, rRNA is maternally inherited and target to rDNA independent of transcription, and de novo transcription is required for proper nucleologenesis in cattle.

AB - 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 µg/ml; total transcriptional inhibition, AD 2.0 µg/ml). Late four-cell embryos were cultured to late eight-cell stage in 0.2 µg/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. RPPI was dispersed. Nuclei (55%) presented rRNA clusters, in 38.8% co-localized with silver-stained deposits. AD 2.0 and ADLT groups displayed no transcription and disintegrating nucleolar precursors. AD 2.0 (34%) and 14% (ADLT) of nuclei presented clusters of maternally inherited rRNA. In AD 2.0 group, RPI was dispersed, but 17.2% of nuclei showed colocalization of rRNA with nucleolar proteins. In ADLT group, RPI was lacking and clustering of nucleolar proteins was hampered. In conclusion, rDNA transcription is not required for targeting of rRNA processing proteins, rRNA is maternally inherited and target to rDNA independent of transcription, and de novo transcription is required for proper nucleologenesis in cattle.

KW - Former LIFE faculty

KW - RNA polymerase I

KW - bovine embryo

KW - genome activation

KW - nucleolus

U2 - 10.1002/mrd.20865

DO - 10.1002/mrd.20865

M3 - Journal article

C2 - 18196555

SN - 1040-452X

VL - 75

SP - 1095

EP - 1103

JO - Molecular Reproduction and Development

JF - Molecular Reproduction and Development

IS - 7

ER -

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

Abstract

Keywords

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