Depletion of BIRC6 leads to retarded bovine early embryonic development and blastocyst formation in vitro
Dessie Salilew-Wondim A , Micheal Hölker A , Franca Rings A , Chirawath Phatsara A , Abdollah Mohammadi-Sangcheshmeh A , Ernst Tholen A , Karl Schellander A and Dawit Tesfaye A B
+ Author Affiliations
- Author Affiliations
A Institute of Animal Science, Animal Breeding and Husbandry Group, University of Bonn, Endenicher Allee 15, Bonn, Germany.
B Corresponding author. Email: tesfaye@itz.uni-bonn.de
Reproduction, Fertility and Development 22(3) 564-579 https://doi.org/10.1071/RD09112
Submitted: 1 May 2009 Accepted: 16 September 2009 Published: 23 February 2010
Abstract
Baculoviral inhibitors of apoptosis repeat-containing 6 (BIRC6) is believed to inhibit apoptosis by targeting key cell-death proteins. To understand its involvement during bovine preimplantation embryo development, two consecutive experiments were conducted by targeted knockdown of its mRNA and protein using RNA interference. In Experiment 1, the effect of BIRC6 knockdown during the early stages of preimplantation embryo development was assessed by injecting zygotes with long double-stranded RNA (ldsRNA) and short hairpin RNA (shRNA) against BIRC6 mRNA followed by in vitro culturing until 96 h post insemination (hpi). The results showed that in RNA-injected zygote groups, reduced levels of BIRC6 mRNA and protein were accompanied by an increase (P < 0.05) in the proportion of 2- and 4-cell and uncleaved embryos and a corresponding decrease (P < 0.05) in the number of 8-cell embryos. In Experiment 2, the effect of BIRC6 knockdown on blastocyst formation, blastocyst total cell number and the extent of apoptosis was investigated. Consequently, zygotes injected with ldsRNA and shRNA resulted in lower (P < 0.05) blastocyst formation and total blastocyst cell number. Moreover, the apoptotic cell ratio, CASPASE 3 and 7 activity, BAX to BCL-2 ratio and levels of SMAC and CASPASE 9 were higher in blastocysts derived from the ldsRNA and shRNA groups, suggesting increased apoptosis in those blastocysts. The results of this study reveal the importance of BIRC6 expression for embryo survival during bovine preimplantation embryo development. However, whether BIRC6 is essential for implantation and fetal development during bovine pregnancy needs further research.
Additional keywords: apoptosis, RNA interference.
References
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