Hum. Reprod. Advance Access originally published online on January 13, 2005
Human Reproduction 2005 20(4):999-1005; doi:10.1093/humrep/deh716
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Exposure of actin on the surface of the human sperm head during in vitro culture relates to sperm morphology, capacitation and zona binding
1 Department of Obstetrics and Gynecology, University of Melbourne, 2 Reproductive Services and 3 Andrology Laboratory, Royal Women's Hospital and 4 Melbourne IVF, Melbourne, Australia
5 To whom correspondence should be addressed. Email: dyl{at}unimelb.edu.au
BACKGROUND: The aim of this study was to determine the relationship between the proportion of motile sperm with actin exposed on the surface of the head and sperm function. METHODS: Semen samples were obtained from normozoospermic men and sperm function tests were performed. Motile sperm selected by swim-up were incubated with actin monoclonal antibody (A-mAb, 1:100) for 2 h, then anti-mouse IgG Dynabeads were used to detect sperm-bound A-mAb. Sperm capacitation was increased by phorbol myristate acetate (PMA) and decreased by bicarbonate-free medium. RESULTS: The proportion of sperm with exposed actin increased with time for up to 2 h incubation. Bicarbonate-free medium significantly decreased the proportion of sperm with exposed actin. PMA significantly enhanced this phenomenon. Sperm bound to zona pellucida (ZP) had a significantly higher proportion with exposed actin than did sperm remaining in medium. Of the 79 samples studied, an average of 9.4% (range 1–27%) of motile sperm had exposed actin after 2 h incubation and this was significantly correlated with sperm normal morphology and ZP binding. CONCLUSION: Exposure of actin on the surface of the sperm head during in vitro culture may be related to membrane modification during sperm capacitation and hence may be a useful marker for this subpopulation of sperm.
Key words: actin/capacitation/morphology motility/sperm–ZP binding
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