Timing and ultrastructure of events following intracytoplasmic sperm injection in a marsupial, the tammar wallaby (Macropus eugenii)
Genevieve M. Magarey A and Karen E. Mate A B C
+ Author Affiliations
- Author Affiliations
A Cooperative Research Centre for Conservation and Management of Marsupials, Macquarie University, NSW 2109, Australia.
B Present address: Division of Biological Sciences, School of Environmental and Life Sciences, University of Newcastle, NSW 2308, Australia.
C To whom correspondence should be addressed. email: karen.mate@newcastle.edu.au
Reproduction, Fertility and Development 15(7) 397-406 https://doi.org/10.1071/RD03033
Submitted: 15 May 2003 Accepted: 25 November 2003 Published: 25 November 2003
Abstract
The aim of the present study was to determine the timing of oocyte activation, sperm decondensation and pronucleus formation after intracytoplasmic sperm injection (ICSI) in the tammar wallaby and to determine the fate of sperm structures at an ultrastructural level. Metaphase II-stage tammar wallaby oocytes were injected with spermatozoa and cultured for 1 (n = 15), 2 (n = 24), 4 (n = 30), 6 (n = 14), 8 (n = 32), 10 (n = 25), 12 (n = 29) or 19 h (n = 12). Oocytes were assessed using light, fluorescence and electron microscopy. The timing of oocyte activation and sperm decondensation after ICSI in the tammar wallaby is relatively similar to that of some eutherian species. Resumption of meiosis II was observed from 1 h and the first female pronucleus was seen 6 h after ICSI. Most oocytes (88%) possessed a female pronucleus by 10 h. Intact acrosomes persisted with intact sperm heads up to 2 h after ICSI. At 10 h, 80% of oocytes possessed a male pronucleus. The sperm tail had undergone considerable degeneration by 10 h after ICSI, including breakdown of the fibrous sheath dense fibres. The identification of sperm tail and midpiece remnants adjacent to pronuclei confirms that the events observed in wallaby oocytes after ICSI are not due to parthenogenetic activation.
Acknowledgments
The authors thank Dr Jim Catt and Sydney IVF (Sydney, Australia) for the loan of micromanipulation equipment, Ms Amanda Harman, Mrs Debra Birch and Ms Janine Buist for skilled technical assistance, Mr Ron Claassens and staff at the Macquarie University Fauna Park for care of the animals and Vetrepharm for the generous donation of pLH. This work was supported by the Australian Government’s Cooperative Research Centres Program. GMM was the recipient of a Macquarie University Postgraduate Research Award.
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