Abstract
Purpose
We investigated the effect of electrical stimulation on rabbit oocyte activation using intracytoplasmic sperm injection (ICSI) to determine whether viable offspring can be produced from deceased rabbit sperm using ICSI.
Methods
Sperm were collected from a heterozygote GFP male rabbit 5 h after sacrifice and cryopreserved in liquid nitrogen. Mature oocytes were fertilized using ICSI. A series of electrical pulse procedures were used to activate oocytes before and/or after ICSI. Following ICSI, zygotes were cultured in B2 medium for 4 days or transferred into the oviducts of recipient rabbits at the 2- or 4-cell stage.
Results
The blastocyst formation rate was significantly greater in oocytes that received one or two pulses prior to ICSI compared to controls and other electrically stimulated groups. In the single pulse before ICSI group, 23 % of the blastocysts expressed GFP, which was significantly greater than all other groups. However, those that received treatment before and after, or just following ICSI, showed a significant decrease in embryo survival. Finally, embryos from the single pulse before ICSI group were transferred into recipient female rabbits and a full-term kit was successfully delivered.
Conclusions
One pulse of electrical stimulation prior to sperm injection was an effective method to activate rabbit oocytes for fertilization. Sperm collected from a deceased rabbit is able to produce viable embryos through ISCI that are capable of normal fetal and kit development.
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Acknowledgments
The study was supported by grants from the Natural Science Foundation of China (Nos. 81170756, 31101048), the Shanghai Natural Science Funding (Nos. 10140901700, 11140901600, 11ZR1418800) and the National Transgenic Foundation of China (2011ZX08008-003).
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Capsule Suitable electrical stimulation on rabbit oocytes could increase the fertilization and embryo development when the sperm from deceased rabbit were used for ICSI, and sperm from deceased rabbit are capable of full term and kit development.
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Zhou, X., Yin, M., Jiang, W. et al. Electrical activation of rabbit oocytes increases fertilization and embryo development by intracytoplasmic sperm injection using sperm from deceased male. J Assist Reprod Genet 30, 1605–1610 (2013). https://doi.org/10.1007/s10815-013-0113-z
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DOI: https://doi.org/10.1007/s10815-013-0113-z