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Viable rabbits derived from oocytes by intracytoplasmic injection of spermatozoa from an infertile male

Published online by Cambridge University Press:  01 May 2009

Qiuyan Li ,
Jian Hou ,
Sheng Wang ,
Hong Guan ,
Nan Zhang ,
Yongfu Chen  and
Xiaorong An
Qiuyan Li
Affiliation:
State Key Laboratory for Agrobiotechnology, College of Biological Sciences, China Agricultural University, Yuan-Ming-Yuan West Road 2, Haidian District, Beijing 100094, China.
Jian Hou
Affiliation:
State Key Laboratory for Agrobiotechnology, College of Biological Sciences, China Agricultural University, Yuan-Ming-Yuan West Road 2, Haidian District, Beijing 100094, China.
Sheng Wang
Affiliation:
State Key Laboratory for Agrobiotechnology, College of Biological Sciences, China Agricultural University, Yuan-Ming-Yuan West Road 2, Haidian District, Beijing 100094, China.
Hong Guan
Affiliation:
State Key Laboratory for Agrobiotechnology, College of Biological Sciences, China Agricultural University, Yuan-Ming-Yuan West Road 2, Haidian District, Beijing 100094, China.
Nan Zhang
Affiliation:
State Key Laboratory for Agrobiotechnology, College of Biological Sciences, China Agricultural University, Yuan-Ming-Yuan West Road 2, Haidian District, Beijing 100094, China.
Yongfu Chen
Affiliation:
State Key Laboratory for Agrobiotechnology, College of Biological Sciences, China Agricultural University, Yuan-Ming-Yuan West Road 2, Haidian District, Beijing 100094, China.
Xiaorong An*
Affiliation:
State Key Laboratory for Agrobiotechnology, College of Biological Sciences, China Agricultural University, Yuan-Ming-Yuan West Road 2, Haidian District, Beijing 100094, China.
*
All correspondence to: Xiaorong An, State Key Laboratory for Agrobiotechnology, College of Biological Sciences, China Agricultural University, Yuan-Ming-Yuan West Road 2, Haidian District, Beijing 100094, China. Tel:/Fax: +86 10 62733463. e-mail: [email protected]
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Summary

Intracytoplasmic sperm injection (ICSI) is an assisted fertilization technique and has been widely applied in human medicine to overcome some obstacles of infertility. However, this technology has not yet been used as a mainstream technique for animal production, including the rabbit, due to its limited success. The aim of this study was to improve ICSI techniques and establish an efficient ICSI method for rabbits. Spermatozoa used for ICSI were collected from mature New Zealand white male rabbits. They were washed two to three times with HEPES-buffered TCM199 containing 10% FBS and then mixed with 10% polyvinylpyrrolidone (PVP) prior to microinjection. Oocytes were harvested from superovulated donor rabbits after 14–15 h hCG treatment and were fertilized by microinjection of a single living spermatozoon into the ooplasm of each oocyte without additional activation treatment. A total of 317 injected oocytes resulted in the high survival rate of 86.1%. Among the surviving oocytes, 273 were placed into culture dishes for in vitro development. The fertilization, cleavage and blastocyst rates were 59.0%, 88.2% and 45.3% respectively. Furthermore, ICSI embryos were produced with spermatozoa from an infertile male rabbit, and 21 early-stage embryos (2-cell and 4-cell) were surgically transferred into the oviducts of two adult female rabbits. On day 31 after transfer, one out of the two recipients gave birth to two normal and healthy young rabbits. These results demonstrate that rabbit oocytes can be successfully activated and fertilized by the new ICSI protocol. Spermatozoa derived from infertile rabbits can successful fertilize oocytes and produce offspring by the simple ICSI technique.

Keywords

Full-term developmentICSIInfertileRabbitSperm
Type
Research Article
Information
Zygote , Volume 17 , Issue 2 , May 2009 , pp. 157 - 162
Copyright
Copyright © Cambridge University Press 2008

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References

Catt, J. & O'Nell, C. (1995). Manipulation of sperm before intracytoplasmic injection improves fertilization rates. Fertil. Steril. 64, 1210–2.CrossRefGoogle ScholarPubMed
Chen, S.H. & Seidel, G.E. Jr. (1997). Effects of oocyte activation and treatment of spermatozoa on embryonic development following intracytoplasmic sperm injection in cattle. Theriogenology 12, 1265–73.CrossRefGoogle Scholar
Chung, J.T., Keefer, L.C. & Downey, B.R. (2000). Activation of bovine oocytes flowing ICSI. Theriogenology 53, 1273–84.CrossRefGoogle Scholar
Dalit, B.Y., Yoger, L., Hauser, R., Haim, Y., Faud, A. & Israel, Y. (1999). Testicular sperm retrieval and cryopreservation prior to initiating ovarian stimulation as the first line approach in patients with non-obstructive azoospermia. Hum. Reprod. 14, 1794–801.Google Scholar
Deng, M. & Yang, X. (2001). Full term development of rabbit oocytes fertilized by intracytoplasmic sperm injection. Mol. Reprod. Dev. 59, 38–43.CrossRefGoogle ScholarPubMed
Gerris, J., Mangelschots, K., Van Royen, E., Joostens, M., Eestermans, W. & Ryckaert, G. (1995). ICSI and severe male-factor infertility: breaking of the sperm tail prior to injection. Hum. Reprod. 10, 484–6.CrossRefGoogle ScholarPubMed
Gomez, M.C., Catt, J.W., Evens, G. & Maxwell, W.M.C. (1998). Cleavage, development and competence of sheep embryos fertilized by ICSI and in vitro fertilization. Theriogenology 49, 1143–54.CrossRefGoogle ScholarPubMed
Hoshi, K., Yazawa, H. & Yanagida, K. (1994). Microinjection of rabbit oocytes with heat- treated sperm: embryonic development. Arch. Androl. 29, 233–7.CrossRefGoogle Scholar
Hosoi, Y. & Iritani, A. (1993). Rabbit microfertilization. Mol. Reprod. Dev. 36, 282–4.CrossRefGoogle ScholarPubMed
Hosoi, Y., Miyake, M., Utsumi, K. & Iritani, A. (1988). Development of rabbit oocytes after microinjection of spermatozoa. In Proceeding of the 11th International Congress on Animal Reproduction and Artificial Insemination. Abstract 331.Google Scholar
Kasai, T., Oshi, K. & Yanagimachi, R. (1999). Effect of sperm immobilization and demembranation on the oocyte activation rate in the mouse. Zygote 7, 187–93.CrossRefGoogle ScholarPubMed
Keefer, C.L. (1989). Fertilization by sperm injection in the rabbit. Gamete Res. 22, 59–69.CrossRefGoogle ScholarPubMed
Kimura, D. & Yanagimachi, R. (1995). Intracytoplasmic sperm injection in mouse. Biol. Reprod. 52, 709–20.CrossRefGoogle Scholar
Kolbe, T. & Holtz, W. (1999). Intracytoplasmic injection (ICSI) of in vivo or in vitro matured oocytes with fresh ejaculated or frozen-thawed epididymal spermatozoa and additional calcium-ionophore activation in the pig. Theriogenology 52, 671–8.CrossRefGoogle ScholarPubMed
Kuretake, S., Kimura, Y., Hoshi, K. & Yanagimachi, R. (1996). Fertilization and development of mouse oocytes injected with isolated sperm heads. Biol. Reprod. 55, 789–95.CrossRefGoogle ScholarPubMed
Li, G.P., Chen, D.Y., Lian, L., Sun, Q.Y., Wang, M.K., Liu, J.L., Li, J.S. & Han, Z.M. (2001). Viable rabbits derived from reconstructed oocytes by germinal vesicle transfer after intracytoplasmic sperm injection (ICSI). Mol. Reprod. Dev. 58, 180–5.3.0.CO;2-7>CrossRefGoogle ScholarPubMed
Li, X.H., Morris, L.H.A. & Allen, W.R. (2000). Effect of different activation treatments on fertilization of horse oocytes by intracytoplasmic sperm injection. J. Reprod. Fertil. 119, 253–60.CrossRefGoogle ScholarPubMed
Liu, J.L., Kusakabe, H., Chang, C.C., Suzuki, H., Schmidt, D., Julian, I., Pfeffer, R. & Charles, L., Bormann, X., Tian, C., Yanagimachi, R. & Yang, X.Z. (2004). Freeze-dried sperm fertilization leads to full-term development in rabbits. Biol. Reprod. 70, 1776–81.CrossRefGoogle ScholarPubMed
Ogonuki, N., Inoue, K., Miki, H., Mochida, K., Hattoti, M., Okada, H., Takeiri, S., Shimozawa, H., Sankai, T. & Ogura, A. (2005). Differential development of rabbit embryos following microinsemination with sperm and spermatids. Mol. Reprod. Dev. 72, 411–7.CrossRefGoogle ScholarPubMed
Palermo, G., Cohn, J. & Rosenwarks, Z. (1996a). Intracytoplasmic sperm injection: a powerful tool to overcome fertilization failure. Fertil. Steril. 65, 899–908.Google ScholarPubMed
Palermo, G., Schlegel, P.N., Colombero, L.T., Zaninovic, N., Moy, F, & Rosenwarks, Z. (1996b). Aggressive sperm immobilization prior to intracytoplasmic sperm injection with immature spermatozoa improves fertilization and pregnancy rates. Hum. Reprod. 11, 1023–9.CrossRefGoogle ScholarPubMed
Rho, G., Kawarsky, S., Johnson, W.H., Kochhar, K. & Betteridge, K.J. (1998a). Sperm and oocyte treatments to improve the formation of male and female pronuclei and subsequent development following intracytoplasmic sperm injection into bovine oocytes. Biol. Reprod. 59, 918–24.CrossRefGoogle ScholarPubMed
Rho, G.J., Wu, B., Kawarsky, S., Leibo, S.P. & Betteridge, K.J. (1998b). Activation regiments to prepare bovine oocytes for intracytoplasmic sperm injection. Mol. Reprod. Dev. 50, 485–92.3.0.CO;2-1>CrossRefGoogle Scholar
Sakamoto, W., Kaneko, T. & Nakagata, N. (2005). Use of frozen-thawed oocytes for efficient production of normal offspring from cryopreserved mouse spermatozoa showing low fertility. Comp. Med. 55, 136–9.Google ScholarPubMed
Sofikitis, N.V., Miyagawa, I., Agapitos, E., Pasyianos, P., Toda, T., Hellstrom, W.J.G. & Kawamura, H. (1994) Reproductive capacity of the nucleus of the male gamete after completion of meiosis. J. Assist. Reprod. Genet. 11, 335–41.CrossRefGoogle ScholarPubMed
Suttner, R., Zakhartchenko, V., Stojkovic, P., Mtillner, S., Alberio, R., Medjugorac, I., Brem, G, Wolf, E. & Stojkovic, M. (2000) Intracytoplasmic sperm injection in bovine: effects of oocyte activation, sperm pretreatment and injection technique. Theriogenology 54, 935–48.CrossRefGoogle ScholarPubMed
Szczygiel, M.A., Kusakabe, H., Yanagimachi, R. & Whittingham, D.G. (2002) Intracytoplasmic sperm injection is more efficient than in vitro fertilization for generating mouse embryos from cryopreserved spermatozoa. Biol. Reprod. 67, 1278–84.CrossRefGoogle ScholarPubMed
Tesarik, J. & Mendoza, C. (2003) Using the male gamete for assisted reproduction: past present and future. J. Androl. 24, 317–28.CrossRefGoogle ScholarPubMed
Tsai, M.Y., Huang, E.L., Kung, E.T., Lin, Y.C., Chang, S.Y, Wu, U. & Chang, H.W. (2000). Influence of polyvinylpyrrolidone on the outcome of intracytoplasmic sperm injection. J. Reprod. Med. 45, 115–22.Google ScholarPubMed
Yavas, Y., Roberge, S., Khamsi, E., Shirazi, P., Endman, M.W, & Wong, J.C. (2001). Performing ICSI using an injection pipette with the smallest possible inner diameter and a long taper increases normal fertilization rate, decreases incidence of degeneration and tripronuclear zygotes, and enhances embryo development. J. Assist. Reprod. Genet. 18, 426–35.CrossRefGoogle Scholar
Zheng, Y.L., Jiang, M.X., Zhang, Y.L., Sun, Q.Y., & Chen, D.Y. (2004). Effect of oocyte age, cumulus cells and injection methods on in vitro development of intracytoplasmic sperm injection in rabbit embryos. Zygote 12, 75–80.CrossRefGoogle ScholarPubMed