Abstract
Objective
The aim of this study was to investigate the developmental competence of oocytes parthenogenetically activated by an electric pulse (EP) and treated with anisomycin and to determine whether this method is applicable to somatic cell nuclear transfer (SCNT).
Results
Embryos derived from porcine oocytes parthenogenetically activated by an EP and treatment with 0.01 µg/mL anisomycin had a significantly improved in vitro developmental capacity. Furthermore, 66.6% of blastocysts derived from these embryos had a diploid karyotype. The blastocyst formation rate of cloned embryos was similar between oocytes activated by an EP and treated with 2 mM 6-dimethylaminopurine for 4 h and those activated by an EP and treated with 0.01 µg/mL anisomycin for 4 h. The level of maturation-promoting factor was significantly decreased in oocytes activated by an EP and treated with anisomycin. Finally, the mRNA expression levels of apoptosis-related genes (Bax and Bcl-2) and pluripotency-related genes (Oct4, Nanog, and Sox2) were checked by RT-PCR.
Conclusion
Our results demonstrate that porcine oocyte activation via an EP in combination with anisomycin treatment can lead to a high blastocyst formation rate in parthenogenetic activation and SCNT experiments.
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Acknowledgements
This work was supported by the State Key Development Program for Basic Research of China (Grant No. 20150622005JC) and the Institute for Basic Science (Grant No. IBS-R021-D1-2015-a02).
Supporting information
Supplementary Table 1—Primer sequences used for gene expression analysis.
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Yu-Chen Zhang, Long Jin, Hai-Ying Zhu have contributed equally to this study.
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Zhang, YC., Jin, L., Zhu, HY. et al. The developmental competence of oocytes parthenogenetically activated by an electric pulse and anisomycin treatment. Biotechnol Lett 39, 189–196 (2017). https://doi.org/10.1007/s10529-016-2249-2
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DOI: https://doi.org/10.1007/s10529-016-2249-2