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Physiology: Physiological Induction of the Acrosome Reaction in Human Sperm: Validation of a Microassay Using Minimal Volumes of Solubilized, Homologous Zona Pellucida

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Abstract

Purpose: The objective was to develop a method that couldaccommodate microvolumes of solubilized human (ZP) andsperm for assessing the induction of the acrosome reaction.

Methods: A microassay using 1 μl of 2.5, 1.25, 0.6, 0.3,and 0.125 ZP/μl incubated with 1 μl of a highly motilesperm suspension for 60 min. As a control and parallelto the microassay a standard acrosome reaction techniquewas performed.

Results: No significant differences were observed betweenthe percentage acrosome-reacted sperm reported by the twoassays under basal conditions (spontaneous) or after inductionwith a Ca2+ ionophore or solubilized ZP. At a ZPconcentration of 0.6 ZP/μl, the percentages ofacrosome-reacted spermatozoa in both techniques were significantlyhigher compared to the spontaneous acrosome reactionresults, namely 18% and 17%, compared to 10% and 10%,respectively. Approximately a 30% level of acrosomal exocytosis was induced with 2.5 ZP/μl in both methods.

Conclusions: This newly devised microtechnique is easy andrapid to perform, is repeatable, and facilitates the use ofminimal volumes of solubilized human ZP (even a single ZP)for assessment of the inducibility of the acrosome reaction ofa homologous sperm population.

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Authors and Affiliations

  1. Department of Obstetrics and Gynaecology, University of Stellen bosch, 7505, Tygerberg, South Africa

    Daniel R. Franken & Hadley S. Bastiaan

  2. Department of Obstetrics and Gynaecology, University of Stellenbosch and Tygerberg Hospital, Tygerberg Hospital, 3rd Floor, 7505, Tygerberg, South Africa

    Daniel R. Franken

  3. Department of Obstetrics and Gynecology, Jones Institute for Reproductive Medicine, Eastern Virginia Medical School, Nor folk, Virginia, 23507

    Sergio C. Oehninger

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  1. Daniel R. Franken
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  2. Hadley S. Bastiaan
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  3. Sergio C. Oehninger
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Franken, D.R., Bastiaan, H.S. & Oehninger, S.C. Physiology: Physiological Induction of the Acrosome Reaction in Human Sperm: Validation of a Microassay Using Minimal Volumes of Solubilized, Homologous Zona Pellucida. J Assist Reprod Genet 17, 156–161 (2000). https://doi.org/10.1023/A:1009418222397

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  • DOI: https://doi.org/10.1023/A:1009418222397

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