Molecular Human Reproduction, Vol. 5, No. 10, 941-949,
October 1999
© 1999 European Society of Human Reproduction and Embryology
Regulation of sperm function |
An important role of actin polymerization in the human zona pellucida-induced acrosome reaction
1 Department of Obstetrics and Gynaecology and 2 Department of Medicine, University of Melbourne, 3 Andrology Laboratory and 4 Reproductive Biology Unit, Royal Women's Hospital, 132 Gratten Street, Carlton, Victoria 3053 and 5 Melbourne IVF, Melbourne, Australia
Abstract
The effects of inhibitors of actin polymerization and depolymerization, cytochalasins and phalloidin, on the human zona pellucida (ZP)-induced acrosome reaction (AR) were investigated. Motile spermatozoa, selected by swim-up technique from normozoospermic men, were incubated in medium with or without the actin modulators. Oocytes (four per test) which had failed to fertilize in vitro were added and incubation continued for 2 h. The spermatozoa bound to the ZP were dislodged by repeatedly aspirating the oocytes with a small-bore pipette and the status of the acrosomes was determined by fluorescein-labelled Pisum sativum agglutinin (PSA). Double immunofluorescent staining with PSA and an anti-actin monoclonal antibody illuminated the acrosomal region of acrosome-intact spermatozoa. In calcium ionophore-induced AR spermatozoa, actin staining was confined to the equatorial segment, post-acrosomal region and tail. Cytochalasins B and D significantly inhibited ZP-induced AR in a dose-dependent manner (P < 0.001). Both inhibitors had no effect on the acrosome of spermatozoa in the insemination medium. Cytochalasin B or D (10–40 µmol/l) had no effect on total percentage motile spermatozoa but decreased sperm velocity and hyperactivation. Phalloidin had no effect on the ZP-induced AR or sperm motility. In conclusion, actin polymerization plays an important role in human ZP-induced AR.
acrosome reaction/actin/cytochalasin B and D/phalloidin
Notes
6 To whom correspondence should be addressed
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