Methods for the Assessment of Sperm Capacitation and Acrosome Reaction Excluding the Sperm Penetration Assay

  • Christopher J. De JongeEmail author
  • Christopher L. R. Barratt
Part of the Methods in Molecular Biology book series (MIMB, volume 927)


Assessing the ability of human spermatozoa to acquire fertilizing potential (capacitation) by stimulating exocytosis of the contents of the acrosome (acrosome reaction) is thought to have diagnostic potential (De Jonge, Reprod Med Rev 3:159–178, 1994). Calcium-mobilizing agents, such as calcium ionophores (A23187) and progesterone, stimulate the acrosome reaction in vitro (Brucker and Lipford, Hum Reprod Update 1:51–62, 1995). Acrosomal status is easily detected using Pisum sativum Agglutinin labeled with fluorescein isothiocyanate (Cross and Meizel, Biol Reprod 41:635–641, 1989). Herein we describe a procedure for assessing capacitation and the acrosome reaction of human spermatozoa in vitro.

Key words

Capacitation Acrosome reaction Calcium ionophore Progesterone PSA-FITC 


  1. 1.
    De Jonge C (2005) Biological basis for human capacitation. Hum Reprod Update 11:205–214PubMedCrossRefGoogle Scholar
  2. 2.
    Ikawa M et al (2010) Fertilization: a sperm’s journey to and interaction with the oocyte. J Clin Invest 120:984–994PubMedCrossRefGoogle Scholar
  3. 3.
    Visconti PE et al (2011) Ion channels, phosphorylation and mammalian sperm capacitation. Asian J Androl 13:395–405PubMedCrossRefGoogle Scholar
  4. 4.
    Zanetti N, Mayorga LS (2009) Acrosomal swelling and membrane docking are required for hybrid vesicle formation during the human sperm acrosome reaction. Biol Reprod 81:396–405PubMedCrossRefGoogle Scholar
  5. 5.
    Yanagimachi R (2011) Mammalian sperm acrosome reaction: where does it begin before fertilization? Biol Reprod 85:4–5PubMedCrossRefGoogle Scholar
  6. 6.
    Visconti PE (2009) Understanding the molecular basis of sperm capacitation through kinase design. Proc Natl Acad Sci U S A 106:667–668PubMedCrossRefGoogle Scholar
  7. 7.
    Cross NL (1996) Human seminal plasma prevents sperm from becoming acrosomally responsive to the agonist, progesterone: cholesterol is the major inhibitor. Biol Reprod 54:138–145PubMedCrossRefGoogle Scholar
  8. 8.
    Cross NL et al (1986) Two simple methods for detecting acrosome-reacted human sperm. Gamete Res 15:213–226CrossRefGoogle Scholar
  9. 9.
    Liu DY, Baker HW (1988) The proportion of human sperm with poor morphology but normal intact acrosomes detected with Pisum sativum agglutinin correlates with fertilization in vitro. Fertil Steril 50:288–293PubMedGoogle Scholar
  10. 10.
    Mendoza C et al (1992) Distinction between true acrosome reaction and degenerative acrosome loss by a one-step staining method using Pisum sativum agglutinin. J Reprod Fertil 95:755–763PubMedCrossRefGoogle Scholar
  11. 11.
    WHO (2010) Laboratory manual for the examination and processing of human semen. World Health Organization, Department of Reproductive Health and Research, GenevaGoogle Scholar
  12. 12.
    Moseley FL et al (2005) Protein tyrosine phosphorylation, hyperactivation and progesterone-induced acrosome reaction are enhanced in IVF media: an effect that is not associated with an increase in protein kinase A activation. Mol Hum Reprod 11:523–529PubMedCrossRefGoogle Scholar
  13. 13.
    Aitken RJ, Brindle JP (1993) Analysis of the ability of three probes targeting the outer acrosomal membrane or acrosomal contents to detect the acrosome reaction in human spermatozoa. Hum Reprod 8:1663–1669PubMedGoogle Scholar
  14. 14.
    Cooper TG, Yeung CH (1998) A flow cytometric technique using peanut agglutinin for evaluating acrosomal loss from human spermatozoa. J Androl 19:542–550PubMedGoogle Scholar
  15. 15.
    Cross NL (1995) Methods for evaluating the acrosomal status of human sperm. In: Fenichel P, Parinaud J (eds) Human sperm acrosome reaction. (Colloques INSERM) John Libbey Eurotext, FranceGoogle Scholar
  16. 16.
    De Jonge CJ et al (1989) Synchronous assay for human sperm capacitation and the acrosome reaction. J Androl 10:232–239PubMedGoogle Scholar
  17. 17.
    Fenichel P et al (1989) Evaluation of the human sperm acrosome reaction using a monoclonal antibody, GB24, and fluorescence-activated cell sorter. J Reprod Fertil 87:699–706PubMedCrossRefGoogle Scholar
  18. 18.
    Henley N et al (1994) Flow cytometric evaluation of the acrosome reaction of human spermatozoa: a new method using a photoactivated supravital stain. Int J Androl 17:78–84PubMedCrossRefGoogle Scholar
  19. 19.
    Talbot P, Chacon RS (1981) A triple-stain technique for evaluating normal acrosome reactions of human sperm. J Exp Zool 215:201–208PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2013

Authors and Affiliations

  • Christopher J. De Jonge
    • 1
    Email author
  • Christopher L. R. Barratt
    • 2
  1. 1.Department of Obstetrics, Gynecology & Women’s HealthUniversity of Minnesota Physicians Reproductive Medicine CenterMinneapolisUSA
  2. 2.Reproductive and Developmental Biology Group, Level 4 MACHS Labs, Division of Medical Sciences, Centre for Oncology and Molecular Medicine, Ninewells HospitalUniversity of DundeeDundeeUK

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