PURPOSE: The objective was to develop a method that could accommodate microvolumes of solubilized human (ZP) and sperm for assessing the induction of the acrosome reaction. METHODS: 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. 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/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. Approximately a 30% level of acrosomal exocytosis was induced with 2.5 ZP/microliter 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.
PURPOSE: The objective was to develop a method that could accommodate microvolumes of solubilized human (ZP) and sperm for assessing the induction of the acrosome reaction. METHODS: 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. 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/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. Approximately a 30% level of acrosomal exocytosis was induced with 2.5 ZP/microliter 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.
Authors: S Oehninger; A A Acosta; L L Veeck; R Brzyski; T F Kruger; S J Muasher; G D Hodgen Journal: Am J Obstet Gynecol Date: 1991-05 Impact factor: 8.661
Authors: D R Franken; W T Oosthuizen; S Cooper; T F Kruger; L J Burkman; C C Coddington; G D Hodgen Journal: Andrologia Date: 1991 May-Jun Impact factor: 2.775