Nuclear and nuclear reprogramming during the first cell cycle in bovine nuclear transfer embryos
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Nuclear and nuclear reprogramming during the first cell cycle in bovine nuclear transfer embryos. / Østrup, Olga; Petrovicova, Ida; Strejcek, Frantisek; Morovic, Martin; Lucas-Hahn, Andrea; Lemme, Erika; Petersen, Bjorn; Niemann, Heiner; Laurincik, Jozef; Hyttel, Poul.
I: Cloning and Stem Cells, Bind 11, Nr. 3, 2009, s. 367-375.Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › fagfællebedømt
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T1 - Nuclear and nuclear reprogramming during the first cell cycle in bovine nuclear transfer embryos
AU - Østrup, Olga
AU - Petrovicova, Ida
AU - Strejcek, Frantisek
AU - Morovic, Martin
AU - Lucas-Hahn, Andrea
AU - Lemme, Erika
AU - Petersen, Bjorn
AU - Niemann, Heiner
AU - Laurincik, Jozef
AU - Hyttel, Poul
PY - 2009
Y1 - 2009
N2 - Abstract The immediate events of genomic reprogramming at somatic cell nuclear transfer (SCNT) are to high degree unknown. This study was designed to evaluate the nuclear and nucleolar changes during the first cell cycle. Bovine SCNT embryos were produced from starved bovine fibroblasts and fixed at 0.5, 1, 2, 3, 4, 8, 12, and 16 h postactivation (hpa). Parthenogenetic (PA) embryos were used as control. The SCNT and PA embryos were processed for lacmoid staining, autoradiography, transmission electron microscopy (TEM), and immunofluorescence localization of: upstream binding factor (UBF) and fibrillarin at 4 and 12 hpa. Likewise, starved and nonstarved fibroblasts were processed for autoradiography and TEM. The fibroblasts displayed strong transcriptional activity and active fibrillogranular nucleoli. None of the reconstructed embryos, however, displayed transcriptional activity. In conclusion, somatic cell nuclei introduced into enucleated oocytes displayed chromatin condensation, partial nuclear envelope breakdown, nucleolar desegregation and transcriptional quiescence already at 0.5 hpa. Somatic cell cytoplasm remained temporally attached to introduced nucleus and nucleolus was partially restored indicating somatic influence in the early SCNT phases. At 1-3 hpa, chromatin gradually decondensed toward the nucleus periphery and nuclear envelope reformed. From 4 hpa, the somatic cell nucleus gained a PN-like appearance and displayed NPBs suggesting ooplasmic control of development.
AB - Abstract The immediate events of genomic reprogramming at somatic cell nuclear transfer (SCNT) are to high degree unknown. This study was designed to evaluate the nuclear and nucleolar changes during the first cell cycle. Bovine SCNT embryos were produced from starved bovine fibroblasts and fixed at 0.5, 1, 2, 3, 4, 8, 12, and 16 h postactivation (hpa). Parthenogenetic (PA) embryos were used as control. The SCNT and PA embryos were processed for lacmoid staining, autoradiography, transmission electron microscopy (TEM), and immunofluorescence localization of: upstream binding factor (UBF) and fibrillarin at 4 and 12 hpa. Likewise, starved and nonstarved fibroblasts were processed for autoradiography and TEM. The fibroblasts displayed strong transcriptional activity and active fibrillogranular nucleoli. None of the reconstructed embryos, however, displayed transcriptional activity. In conclusion, somatic cell nuclei introduced into enucleated oocytes displayed chromatin condensation, partial nuclear envelope breakdown, nucleolar desegregation and transcriptional quiescence already at 0.5 hpa. Somatic cell cytoplasm remained temporally attached to introduced nucleus and nucleolus was partially restored indicating somatic influence in the early SCNT phases. At 1-3 hpa, chromatin gradually decondensed toward the nucleus periphery and nuclear envelope reformed. From 4 hpa, the somatic cell nucleus gained a PN-like appearance and displayed NPBs suggesting ooplasmic control of development.
U2 - 10.1089/clo.2008.0076
DO - 10.1089/clo.2008.0076
M3 - Journal article
C2 - 19594386
VL - 11
SP - 367
EP - 375
JO - Cellular Reprogramming
JF - Cellular Reprogramming
SN - 2152-4971
IS - 3
ER -
ID: 14306955