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Effects of sucrose treatment on the development of mouse nuclear transfer embryos with morula blastomeres as donors

Published online by Cambridge University Press:  01 February 2008

Gang Zhang ,
Qing-Yuan Sun  and
Da-Yuan Chen
Gang Zhang
Affiliation:
State Key Laboratory of Reproductive Biology, Institute of Zoology, The Chinese Academy of Sciences, Beijing, 100101, China. Department of Cell and Systems Biology, University of Toronto, 25 Harbord Street, RW618, Toronto, Ontario, M5S 3G8, Canada.
Qing-Yuan Sun
Affiliation:
State Key Laboratory of Reproductive Biology, Institute of Zoology, The Chinese Academy of Sciences, Beijing, 100101, China.
Da-Yuan Chen*
Affiliation:
State Key Laboratory of Reproductive Biology, Institute of Zoology, The Chinese Academy of Sciences, Beijing, 100101, China.
*
1All correspondence to: Da-Yuan Chen. State Key Laboratory of Reproductive Biology, Institute of Zoology, The Chinese Academy of Sciences, Datun Road, Chaoyang District, Beijing, 100101, China. Tel: +86 010 64807052. Fax: +86 010 64807316. e-mail: [email protected]
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Summary

In this study, nuclear transfer (NT) embryos were produced by using C57Bl/6 mouse morula blastomeres and Kunming mouse metaphase II (MII) oocytes as donors and recipients, respectively, to investigate the effects of sucrose treatment of MII oocytes with different concentrations on the manipulation time of NT, electrofusion and the in vitro and in vivo development of reconstructed embryos. The results demonstrated that: (i) when the oocytes were enucleated with 1, 2 and 3% sucrose treatment, respectively, the enucleating rates were not affected by the different sucrose concentrations, but the manipulation time had significant difference and the mean nuclear transfer manipulation times of every oocyte were 180 ± 10 s, 130 ± 10 s and 120 ± 10 s, respectively; (ii) different sucrose concentrations had no significant effects on the fusion rate and the in vitro developmental potential of the NT embryos (p > 0.05). Furthermore, 59 embryos were transplanted into the oviducts of two recipients. In the end, three dead full-term developed fetuses were obtained on 21 days post coitus (dpc). These results suggested that the mouse MII oocytes enucleated via sucrose treatment might be an alternative source for mouse cloning and could support the embryonic NT embryos developed to term in vivo.

Keywords

Morula blastomeresMouseNuclear transferSucrose treatment
Type
Research Article
Information
Zygote , Volume 16 , Issue 1 , February 2008 , pp. 15 - 19
Copyright
Copyright © Cambridge University Press 2008

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References

Cheong, H.T., Takahashi, Y. & Kanagawa, H. (1993). Birth of mice after transplantation of early cell-cycle-stage embryonic nuclei into enucleated oocytes. Biol. Reprod. 48, 958–63.CrossRefGoogle ScholarPubMed
Eggan, K., Baldwin, K., Tackett, M., Osborne, J., Gogos, J., Chess, A., Axel, R. & Jaenisch, R. (2004). Mice cloned from olfactory sensory neurons. Nature 428, 44–9.CrossRefGoogle ScholarPubMed
Hochedlinger, K. & Jaenisch, R. (2002). Monoclonal mice generated by nuclear transfer from mature B and T donor cells. Nature 415, 1035–8.CrossRefGoogle ScholarPubMed
Kong, F.Y., Zhang, G., Zhong, Z.S., Li, Y.L., Sun, Q.Y. & Chen, D.Y. (2005). Transplantation of male pronucleus derived from in vitro fertilization of enucleated oocyte into parthenogenetically activated oocyte results in live offspring in mouse. Zygote 13, 35–8.CrossRefGoogle Scholar
Kono, T., Kwon, O.Y. & Nakahara, T. (1991). Development of enucleated mouse oocytes reconstituted with embryonic nuclei. J. Reprod. Fert. 93, 165–72.CrossRefGoogle ScholarPubMed
Li, J., Ishii, T., Feinstein, P. & Mombaerts, P. (2004). Odorant receptor gene choice is reset by nuclear transfer from mouse olfactory sensory neurons. Nature 428, 393–9.CrossRefGoogle ScholarPubMed
McGrath, J. & Solter, D. (1983). Nuclear transplantation in the mouse embryo by microsurgery and cell fusion. Science 220, 1300–2.CrossRefGoogle ScholarPubMed
Mohamed Nour, M.S. & Takahashi, Y. (1999). Preparation of young preactivated oocytes with high enucleation efficiency for bovine nuclear transfer. Theriogenology 51, 661–6.CrossRefGoogle Scholar
Ogura, A., Inoue, K., Ogonuki, N., Noguchi, A., Takano, K., Nagano, R., Suzuki, O., Lee, J., Ishino, F. & Matsuda, J. (2000a). Production of male cloned mice from fresh, cultured and cryopreserved immature Sertoli cells. Biol. Reprod. 62, 1579–84.CrossRefGoogle ScholarPubMed
Ogura, A., Inoue, K., Takano, K., Wakayama, T. & Yanagimachi, R. (2000b). Birth of mice after nuclear transfer by electrofusion using tail tip cells. Mol. Reprod. Dev. 57, 55–9.3.0.CO;2-W>CrossRefGoogle ScholarPubMed
Ono, Y., Shimozawa, N., Ito, M. & Kono, T. (2001). Cloned mice from fetal fibroblast cells arrested at metaphase by serial nuclear transfer. Biol. Reprod. 64, 44–50.CrossRefGoogle ScholarPubMed
Tsunoda, Y. & Kato, Y. (1997). Full-term development after transfer of nuclei from 4-cell and compacted morula stage embryos to enucleated oocytes in the mouse. J. Exp. Zool. 278, 250–4.3.0.CO;2-O>CrossRefGoogle ScholarPubMed
Tsunoda, Y. & Kato, Y. (1998). Not only inner cell mass cell nuclei but also trophectoderm nuclei of mouse blastocysts have a developmental totipotency. J. Reprod. Fertil. 113, 181–4.CrossRefGoogle ScholarPubMed
Tsunoda, Y., Yasui, T., Nakamura, K., Uchida, T. & Sugie, T. (1986). Effect of cutting the zona pellucida on the pronuclear transplantation in the mouse. J. Exp. Zool. 240, 119–25.CrossRefGoogle ScholarPubMed
Tsunoda, Y., Shioda, Y., Onodera, M., Nakamura, K. & Uchida, T. (1988). Differential sensitivity of mouse pronuclei and zygote cytoplasm to Hoechst staining and ultraviolet irradiation. J. Reprod. Fertil. 82, 173–8.CrossRefGoogle ScholarPubMed
Wakayama, T., Perry, A.C., Zuccotti, M., Johnson, K.R. & Yanagimachi, R. (1998). Full-term development of mice from enucleated oocytes injected with cumulus cell nuclei. Nature 394, 369–74.CrossRefGoogle ScholarPubMed
Wakayama, T., Rodriguez, I., Perry, A.C., Yanagimachi, R. & Mombaerts, P. (1999a). Mice cloned from embryonic stem cells. Proc. Natl. Acad. Sci. USA. 96, 14984–9.CrossRefGoogle ScholarPubMed
Wakayama, T. & Yanagimachi, R. (1999b). Cloning of male mice from adult tail-tip cells. Nat. Genet. 22, 127–8.CrossRefGoogle ScholarPubMed
Wang, M.K., Chen, D.Y., Liu, J.L., Li, G.P. & Sun, Q.Y. (2001a). In vitro fertilization of mouse oocytes reconstructed by transfer of metaphase 2 chromosomes results in live births. Zygote 9, 9–14.CrossRefGoogle Scholar
Wang, M.K., Liu, J.L., Li, G.P., Lian, L. & Chen, D.Y. (2001b). Sucrose pretreatment for enucleation: an efficient and non-damage method for removing the spindle of the mouse M2 oocytes. Mol. Reprod. Dev. 58, 432–6.3.0.CO;2-Y>CrossRefGoogle Scholar