Silencing CENPF in bovine preimplantation embryo induces arrest at 8-cell stage

• Published in: Reproduction (Cambridge, England) Vol. 138; no. 5; pp. 783 - 791
• Main Authors: Toralová, Tereza, Šušor, Andrej, Němcová, Lucie, Kepková, Kateřina, Kaňka, Jiří
• Format: Journal Article
• Language: English
• Published: England BioScientifica 01.11.2009; BioScientifica Ltd; Society for Reproduction and Fertility
• Subjects: Animals; Blastocyst - cytology; Blastocyst - metabolism; Blastocyst - physiology; Cattle - embryology; Cattle - genetics; Cattle - physiology; Cell Cycle Checkpoints - genetics; Cell Cycle Checkpoints - physiology; Cell Division - genetics; Cell Division - physiology; Cells, Cultured; Chromosomal Proteins, Non-Histone - antagonists & inhibitors; Chromosomal Proteins, Non-Histone - genetics; Chromosomal Proteins, Non-Histone - metabolism; Cleavage Stage, Ovum - metabolism; Cleavage Stage, Ovum - physiology; Embryonic Development - genetics; Embryonic Development - physiology; Female; Fertilization in Vitro - veterinary; Gene Expression Regulation, Developmental; Gene Silencing - physiology; Microfilament Proteins - antagonists & inhibitors; Microfilament Proteins - genetics; Microfilament Proteins - metabolism
• ISSN: 1470-1626; 1741-7899
• DOI: 10.1530/REP-09-0234

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.