Molecular Human Reproduction, Vol. 8, No. 1, 37-47,
January 2002
© 2002 European Society of Human Reproduction and Embryology
Testis and spermatogenesis |
An anti-actin monoclonal antibody inhibits the zona pellucida-induced acrosome reaction and hyperactivated motility of human sperm
1 Department of Obstetrics and Gynecology, 4 Anatomy and Cell Biology and 5 Medicine, University of Melbourne, 2 Reproductive Services and 3 Andrology Laboratory, Royal Women's Hospital and 6 Melbourne IVF, Melbourne, Australia
We report an inhibitory effect of an anti-actin monoclonal antibody (mAb) on the human zona pellucida (ZP)-induced acrosome reaction (AR). Motile sperm were incubated with native human ZP for 2 h in medium containing either the anti-actin mAb, an irrelevant control mAb or cytochalasins B or D (40 µmol/l). Sperm bound to the ZP were recovered and the AR was determined by fluorescein-labelled Pisum Sativum agglutinin. Anti-mouse immunoglobulin G (mIgG) Dynabeads, immunofluorescence and immunogold were used to detect the location of the anti-actin mAb in sperm. The anti-actin mAb significantly inhibited the ZP-induced AR (equivalent to cytochalasins), the ionophore A23187-induced AR and hyperactivation of sperm in medium. After incubation with anti-actin mAb, anti-mIgG beads bound to the head of >50% of sperm recovered after binding to the ZP and 10% of sperm remaining in the medium. The proportion of sperm that bound anti-mIgG beads after recovery from binding to the ZP in the presence of the anti-actin mAb was significantly correlated with the ZP-induced AR in the absence of the antibody. Immunofluorescence and immunogold demonstrated entry of the anti-actin mAb into sperm. This study suggests that the sperm plasma membrane becomes permeable to the anti-actin mAb during capacitation and initiation of the AR.
actin/actin monoclonal antibody/cytochalasin/human acrosome reaction/sperm
7 To whom correspondence should be addressed at: Department of Obstetrics and Gynecology, University of Melbourne, Royal Women's Hospital, 132 Grattan St, Carlton, Victoria 3053, Australia. E-mail: dyl{at}unimelb.edu.au
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