Prolonged interval between fusion and activation impairs embryonic development by inducing chromosome scattering and nuclear aneuploidy in pig somatic cell nuclear transfer
Jinyoung You A D , Kilyoung Song B D and Eunsong Lee A C E
+ Author Affiliations
- Author Affiliations
A College of Veterinary Medicine, Kangwon National University, Chuncheon 200-701, Korea.
B JCOM R&D Center, Daedae-ri 289, Cheoin-gu, Yongin 449-822, Korea.
C Institute of Veterinary Science, Kangwon National University, Chuncheon 200-701, Korea.
D These authors contributed equally to this work.
E Corresponding author. Email: eslee@kangwon.ac.kr
Reproduction, Fertility and Development 22(6) 977-986 https://doi.org/10.1071/RD09309
Submitted: 18 December 2009 Accepted: 27 January 2010 Published: 1 July 2010
Abstract
The aim of the present study was to examine the effect of various intervals between electrofusion and activation (FA interval) on the nuclear remodelling and development of somatic cell nuclear transfer (SCNT) embryos in pigs. Reconstructed oocytes were activated at 0 (simultaneous fusion and activation; SFA), 1, 2 and 3 h (delayed activation) after electrofusion; these groups were designated as DA1, DA2 and DA3, respectively. When oocyte nuclear status was examined at 0.5, 1, 2 and 3 h after electrofusion, the incidence of chromosome scattering was increased (P < 0.01) as the FA interval was extended (0.0%, 12.0%, 77.3% and 78.0%, respectively). Extending the FA interval led to an increase (P < 0.01) in the percentage of oocytes containing multiple (≥3) pseudopronuclei (PPN) (0.0% of SFA; 5.3% of DA1; 21.7% of DA2; and 33.5% of DA3). The development of SCNT embryos to the blastocyst stage was decreased (P < 0.05) in DA2 (5.7%) and DA3 (5.0%) compared with SFA (18.1%) and DA1 (19.5%). Our results demonstrate that extending the FA interval impairs the development of SCNT pig embryos by inducing chromosome scattering and the formation of multiple PPN, which may result in increased nuclear aneuploidy.
Additional keywords: maturation promoting factor, nuclear ploidy, nuclear remodelling, pseudopronucleus.
Acknowledgements
The authors thank Gyeonggi Veterinary Service (Suwon, Gyeonggi-do, Korea) for supplying pig ovaries and Ms Eunjin Kwon for transporting the pig ovaries. This research was supported by Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education, Science and Technology (2009-0072438).
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