Synchronous assay for human sperm capacitation and the acrosome reaction

Journal of Andrology
C De JongeL J Zaneveld

Abstract

A synchronous acrosome reaction system was established for human spermatozoa. Seminal plasma is removed from the spermatozoa by centrifugation and the washed spermatozoa are capacitated in a modified BWW medium (without exogenous substrates) containing 35 mg/ml human serum albumin for 3 h at 37 C. Subsequently, 10 microM ionophore A23187 (final concentration) is added, the mixture incubated for 15 min at 37 C and the percent acrosome reaction determined by a modified triple stain technique (trypan blue stain omitted). Since no significant decrease in sperm motility occurs during incubation or after ionophore treatment, a vital stain does not need to be employed, allowing the use of any acrosome detection technique. The average percentage of acrosome-rejected spermatozoa after ionophore treatment (40 +/- 10%) was about 2- to 3-fold higher than that seen without ionophore treatment. Ionophore treatment for 15 min failed to stimulate the acrosome reaction in spermatozoa incubated for less than 3 h. Additionally, the presence of substrates in the BWW medium, higher sperm numbers, increased ionophore concentrations or longer incubation periods did not enhance the induction of the acrosome reaction. Ionomycin, a more specific calcium...Continue Reading

References

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Citations

Feb 1, 1992·Journal of Assisted Reproduction and Genetics·R S JeyendranA C Wentz
Mar 1, 1995·Molecular Reproduction and Development·R A AndersonL J Zaneveld
Feb 1, 1995·Molecular Reproduction and Development·P J AshworthP F Watson
Nov 1, 1993·Molecular Reproduction and Development·H L Boettger-TongG R Poirier
Apr 1, 1997·American Journal of Reproductive Immunology : AJRI·S DorjeeC Y Lee
Jan 25, 2017·Human Reproduction Update·Christopher De Jonge
Dec 15, 1992·The Journal of Experimental Zoology·C A Holden, A O Trounson
Oct 20, 1999·Andrologia·C A Villanueva-DiazS Karchmer-Krivitsky
Aug 2, 2001·The Journal of Experimental Zoology·A MinelliC Castellini
Jun 1, 1993·Molecular Reproduction and Development·R A HarrisonN G Miller
Jan 6, 2018·Human Reproduction·Dorte Louise Egeberg PalmeKristian Almstrup
Aug 1, 1989·Gamete Research·C J De JongeL J Zaneveld
Nov 1, 1992·The Journal of Experimental Zoology·P F WatsonP S Jones

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