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Mouse sperm fertilising capacity following subzonal microinjection is dependent on sperm washing and response to solubilised zonae pellucidae

Published online by Cambridge University Press:  26 September 2008

Orly Lacham Kaplan  and
Alan Trounson
Orly Lacham Kaplan
Affiliation:
Institute for Reproduction and Development, Monash University, Melbourne, Australia.
Alan Trounson*
Affiliation:
Institute for Reproduction and Development, Monash University, Melbourne, Australia.
*
Dr Alan Trounson, Centre for Early Human Development, Monash University, Monash Medical Centre, 246 Clayton Road, Clayton, Victoria 3168, Australia. Telephone: 61-3-550-5300. Fax: 61-3-550 5554.
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Summary

Spermatozoa obtained from hybrid male mice were treated with solubilised zonae pellucidae after a period of capacitation in vitro to induce the acrosome reaction. Single spermatozoa were selected and microinjected into the perivitelline space of mature oocytes. A high proportion of the spermatozoa acrosome-reacted affter treatment with solubilised zonae (63%). However, the fertilisation rate (37%) after subzonal microinjection of oocytes by the treated spermatozoa was not different to the fertilisation rate (40%) of oocytes microinjected with untreated spermatozoa which had a lower rate of acrosome reaction (39%). When spermatozoa were washed by high-speed centrifugation before treatment with solubilised zonae pellucidae and subzonal microinjection, the fertilisation rate (68%) was significantly higher (p < 0.001) than that (29%) for occytes microinjected with untreated spermatozoa and was found to be correlated with a high acrosome reaction rate (74%) (r = 0.8). The washing of spermatozoa by centrifugation itself did not increase the acrosome reaction rate or fertilisation rate of oocytes after microinjection. The results of this study suggests that some modifications other than the acrosome reaction are needed to enable capacitated mouse spermatozoa to fuse with the oocyte plasma membrane. These modifications were achieved by washing spermatozoa by high-speed centrifugation and the replacement of the supernatant with fresh culture medium used for capacitation. Induction of the acrosome reaction by solubilised zonae pellucidae following this treatment leads to a high fertilisation rate of oocytes by subzonal sperm microinjection.

Keywords

Acrosome reactionFertilisationMicroinjectionSpermatozoaZona pellucida
Type
Article
Information
Zygote , Volume 3 , Issue 1 , February 1995 , pp. 9 - 16
Copyright
Copyright © Cambridge University Press 1995

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