Silencing CENPF in bovine preimplantation embryo induces arrest at 8-cell stage.
Identification of genes that are important for normal preimplantation development is essential for understanding the basics of early mammalian embryogenesis. In our previous study, we have shown that CENPF (mitosin) is differentially expressed during preimplantation development of bovine embryos. CENPF is a centromere-kinetochore complex protein that plays a crucial role in the cell division of somatic cells. To our best knowledge, no study has yet been done on either bovine model, or oocytes and preimplantation embryos. In this study, we focused on the fate of bovine embryos after injection of CENPF double-stranded RNA (dsRNA) into the zygotes. An average decrease of CENPF mRNA abundance by 94.9% or more and an extensive decline in immunofluorescence staining intensity was detected relative to controls. There was no disparity between individual groups in the developmental competence before the 8-cell stage. However, the developmental competence rapidly decreased then and only 28.1% of CENPF dsRNA injected 8-cell embryos were able to develop further (uninjected control: 71.8%; green fluorescent protein dsRNA injected control: 72.0%). In conclusion, these results show that depletion of CENPF mRNA in preimplantation bovine embryos leads to dramatic decrease of developmental competence after embryonic genome activation.
Toralová
Tereza
T
Department of Reproductive and Developmental Biology, Institute of Animal Physiology and Genetics, vvi, Academy of Sciences of the Czech Republic, Libechov, Czech Republic. moravcova@iapg.cas.cz
Susor
Andrej
A
Nemcová
Lucie
L
Kepková
Katerina
K
Kanka
Jirí
J
D000818:Animals; D001755:Blastocyst / Q000166:cytology* / Q000378:metabolism* / Q000502:physiology; D002417:Cattle / Q000196:embryology / Q000235:genetics / Q000502:physiology; D059447:Cell Cycle Checkpoints / Q000235:genetics / Q000502:physiology; D002455:Cell Division / Q000235:genetics* / Q000502:physiology; D002478:Cells, Cultured; D002868:Chromosomal Proteins, Non-Histone / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000378:metabolism; D002970:Cleavage Stage, Ovum / Q000378:metabolism / Q000502:physiology; D047108:Embryonic Development / Q000235:genetics / Q000502:physiology; D005260:Female; D005307:Fertilization in Vitro / Q000662:veterinary; D018507:Gene Expression Regulation, Developmental; D020868:Gene Silencing / Q000502:physiology*; D008840:Microfilament Proteins / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000378:metabolism
D002868:Chromosomal Proteins, Non-Histone; D008840:Microfilament Proteins; C094772:centromere protein F