Physiological induction of the acrosome reaction in human sperm: Validation of a microassay using minimal volumes of solubilized, homologous zona pellucida
Date
2000
Authors
Franken D.R.
Bastiaan H.S.
Oehninger S.C.
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Abstract
Purpose: To develop a method that could accommodate microvolumes of solubilized human zona pellucida (ZP) and sperm 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 motile sperm suspension for 60 min. As a control and parallel to the microassay a standard acrosome reaction technique was performed. Results: No significant differences were observed between the percentage acrosome reacted sperm reported by the two assays under basal conditions (spontaneous) or after induction with a Ca2+ ionophore or solubilized ZP. At a ZP concentration of 0.6 ZP/μl, the percentages of acrosome-reacted spermatozoa in both techniques were significantly higher compared to the spontaneous acrosome reaction results, namely, 18% and 17%, compared to 10% and 10%, respectively. An approximately 30% level of acrosomal exocytosis was induced with 2.5 ZP/μl in both methods. Conclusions: This newly devised microtechnique is easy and rapid to perform, is repeatable and facilitates the use of minimal volumes of solubilized human ZP (even a single ZP) for assessment of the inducibility of the acrosome reaction of a homologous sperm population.
Description
Keywords
calcium ionophore, acrosome reaction, analytic method, article, assay, controlled study, exocytosis, human, human cell, intermethod comparison, male, priority journal, solubilization, sperm, validation process, zona pellucida, Acrosome Reaction, Biological Assay, Calcium, Female, Human, Ionophores, Male, Solubility, Spermatozoa, Zona Pellucida
Citation
Journal of Assisted Reproduction and Genetics
17
7
17
7