Physiological induction of the acrosome reaction in human sperm : Validation of a microassay using minimal volumes of solubilized, homologous zona pellucida

To develop a method that could accommodate microvolumes of solubilized human zona pellucida (ZP) and sperm for assessing the induction of the acrosome reaction. A microassay using 1 microliter of 2.5, 1.25, 0.6, 0.3, and 0.125 ZP/microliter incubated with 1 microliter of a highly motile sperm suspen...

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Published in:Journal of assisted reproduction and genetics 2000-08, Vol.17 (7), p.374-378
Main Authors: FRANKEN, Daniel R, BASTIAAN, Hadley S, OEHNINGER, Sergio C
Format: Article
Language:English
Subjects:
Acrosome Reaction - drug effects
Acrosome Reaction - physiology
Biological and medical sciences
Biological Assay - methods
Calcium - chemistry
Calcium - metabolism
Female
Fundamental and applied biological sciences. Psychology
Humans
In vitro fertilization
Ionophores - pharmacology
Laboratories
Male
Mammalian reproduction. General aspects
Motility
Physiology
Solubility
Sperm
Spermatozoa - drug effects
Spermatozoa - physiology
Vertebrates: reproduction
Zona Pellucida - chemistry
Zona Pellucida - physiology
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Summary:To develop a method that could accommodate microvolumes of solubilized human zona pellucida (ZP) and sperm for assessing the induction of the acrosome reaction. A microassay using 1 microliter of 2.5, 1.25, 0.6, 0.3, and 0.125 ZP/microliter incubated with 1 microliter of a highly motile sperm suspension for 60 min. As a control and parallel to the microassay a standard acrosome reaction technique was performed. 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/microliter, 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/microliter in both methods. 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.
ISSN:1058-0468
1573-7330
DOI:10.1023/A:1009493708268