Nucleolar development and allocation of key nucleolar proteins require de novo transcription in bovine embryos
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Nucleolar development and allocation of key nucleolar proteins require de novo transcription in bovine embryos. / Svarcova, Olga; Laurincik, Jozef; Avery, Birthe; Mlyncek, Milos; Niemann, Heiner; Maddox-Hyttel, Poul.
I: Molecular Reproduction and Development, Bind 74, Nr. 11, 2007, s. 1428-1435.Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › fagfællebedømt
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TY - JOUR
T1 - Nucleolar development and allocation of key nucleolar proteins require de novo transcription in bovine embryos
AU - Svarcova, Olga
AU - Laurincik, Jozef
AU - Avery, Birthe
AU - Mlyncek, Milos
AU - Niemann, Heiner
AU - Maddox-Hyttel, Poul
PY - 2007
Y1 - 2007
N2 - The goal of the present study was to investigate whether key nucleolar proteins involved in ribosomal RNA (rRNA) transcription and processing are transcribed de novo or from maternally inherited messenger RNAs (mRNA) in bovine embryos, and to which extent de novo transcription of these proteins mRNA is required for the development of functional nucleoli during the major activation of the embryonic genome. Immunofluorescence for localization of key nucleolar proteins, autoradiography for detection of transcriptional activity, and transmission electron microscopy were applied to in vitro produc ed bovine embryos cultured from the 2-cell stage with or without (control groups) a-amanitin, which blocks the RNA plymerases II and III transcription and, thus the synthesis of mRNA. In the control groups, weak autoradiographic labelling was initially observed in the periphery of few nuclei at the 4-cell and the early 8-cell stage, and the entire nucleoplasm as well as nucleolus precursor bodies (NBBs) were prominently labelled in all late 8-cell stages. The NPBs displayed initial transformation into fibrillo-granular nucleoli. In the a-amanitin group, lack of autoradiographic labelling was seen at all developmental stages and disintegrated NPBs stage were found at the late 8-cell. Our immunofluorescence data indicate that RNA polymerase I, UBF, topoisomerase I and fibrillarin are transcribed de novo whereas nucleolin and nucleophosmin are maternally inherited as demonstrated by a-amanitin inhibition. However, localization of these two proteins to the nucleolar compartments was negatively affected by the a-aminitin treatment. Consequentl, functional nucleoli were not established.
AB - The goal of the present study was to investigate whether key nucleolar proteins involved in ribosomal RNA (rRNA) transcription and processing are transcribed de novo or from maternally inherited messenger RNAs (mRNA) in bovine embryos, and to which extent de novo transcription of these proteins mRNA is required for the development of functional nucleoli during the major activation of the embryonic genome. Immunofluorescence for localization of key nucleolar proteins, autoradiography for detection of transcriptional activity, and transmission electron microscopy were applied to in vitro produc ed bovine embryos cultured from the 2-cell stage with or without (control groups) a-amanitin, which blocks the RNA plymerases II and III transcription and, thus the synthesis of mRNA. In the control groups, weak autoradiographic labelling was initially observed in the periphery of few nuclei at the 4-cell and the early 8-cell stage, and the entire nucleoplasm as well as nucleolus precursor bodies (NBBs) were prominently labelled in all late 8-cell stages. The NPBs displayed initial transformation into fibrillo-granular nucleoli. In the a-amanitin group, lack of autoradiographic labelling was seen at all developmental stages and disintegrated NPBs stage were found at the late 8-cell. Our immunofluorescence data indicate that RNA polymerase I, UBF, topoisomerase I and fibrillarin are transcribed de novo whereas nucleolin and nucleophosmin are maternally inherited as demonstrated by a-amanitin inhibition. However, localization of these two proteins to the nucleolar compartments was negatively affected by the a-aminitin treatment. Consequentl, functional nucleoli were not established.
KW - Former LIFE faculty
KW - embryo
KW - maternal-embryonic transition
KW - nucleolus
KW - gene expression
KW - alpha-amanitin
U2 - 10.1002/mrd.20727
DO - 10.1002/mrd.20727
M3 - Journal article
C2 - 17410544
VL - 74
SP - 1428
EP - 1435
JO - Molecular Reproduction and Development
JF - Molecular Reproduction and Development
SN - 1040-452X
IS - 11
ER -
ID: 8078378