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    López-Saucedo, J. Paramio-Nieto, M.T. Fierro, R. and Piña-Aguilar, R.E. 2012. Intracytoplasmic sperm injection (ICSI) in small ruminants. Animal Reproduction Science, Vol. 133, Issue. 3-4, p. 129.


    Rahman, Abu Nasar Md. Aminoo 2010. Intracytoplasmic Sperm Injection-Revolution in Human and Animal Assisted Reproduction: A Review. Biotechnology(Faisalabad), Vol. 9, Issue. 4, p. 392.


    Nation, A and Selwood, L 2009. The production of mature oocytes from adult ovaries following primary follicle culture in a marsupial. Reproduction, Vol. 138, Issue. 2, p. 247.


    Czarny, N. A. Mate, K. E. and Rodger, J. C. 2008. Acrosome stability in the spermatozoa of dasyurid marsupials. Reproduction, Fertility and Development, Vol. 20, Issue. 2, p. 295.


    Richings, Nadine M. Shaw, Geoffrey Temple-Smith, Peter D. and Renfree, Marilyn B. 2006. Growth and histology of ovarian follicles after cold storage in the tammar wallaby. Reproduction, Fertility and Development, Vol. 18, Issue. 6, p. 677.


    Richings, N. M Shaw, G. Temple-Smith, P. D and Renfree, M. B 2004. Intra-cytoplasmic sperm injection in a marsupial. Reproduction, Vol. 128, Issue. 5, p. 595.


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Fertilization following intracytoplasmic sperm injection of in vivo and in vitro matured oocytes from an Australian marsupial, the tammar wallaby (Macropus eugenii)

  • Genevieve M. Magarey (a1) and Karen E. Mate (a1)
  • DOI: http://dx.doi.org/10.1017/S0967199403002430
  • Published online: 01 March 2004
Abstract

Conventional in vitro fertilization (IVF) techniques have been unable to produce normal embryos in any Australian marsupial, largely owing to problems with the early stages of sperm–oocyte binding. This study has used intracytoplasmic sperm injection (ICSI) of in vivo and in vitro matured tammar wallaby oocytes to bypass these processes and achieve fertilization in vitro. The fertilization rate (i.e. development to the two-pronuclei stage) of in vivo and in vitro matured oocytes following ICSI and sham injection was assessed at 17–19 h after injection. Fertilization occurred in 48% (45/93) of in vivo matured oocytes that were injected with spermatozoa. Significantly fewer sham-injected oocytes (6/82, P<0.005) and uninjected control oocytes (5/84, P<0.005) formed two pronuclei. In a direct comparison, the numbers of in vivo and in vitro matured oocytes that formed two pronuclei after ICSI were 22/28 (78.6%) and 23/40 (57.6%), respectively, which are not significantly different. There was also no significant difference in the nuclear response of in vivo and in vitro matured oocytes to sham injection. The numbers of oocytes forming a single pronucleus after sham injection were 10/24 (41.7%) and 24/37 (64.9) for in vivo and in vitro matured oocytes, respectively. Immature germinal-vesicle-stage oocytes were unable to decondense sperm injected during ICSI or to form pronuclei. These results demonstrate that both in vitro and in vivo matured tammar wallaby oocytes can be fertilized by ICSI. The success of ICSI not only offers the opportunity for fundamental analysis of marsupial fertilization but could, in conjunction with development of appropriate culture conditions and embryo transfer technologies, contribute to increased production of offspring from rare or valuable marsupials.

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Copyright
Corresponding author
All correspondence to: K. Mate, School of Environmental and Life Sciences, University of Newcastle, NSW 2308, Australia. Tel: +61 2 49217102. Fax: +61 2 49216899. e-mail: Karen.Mate@newcastle.edu.au
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Zygote
  • ISSN: 0967-1994
  • EISSN: 1469-8730
  • URL: /core/journals/zygote
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