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Hum. Reprod. Advance Access published online on July 24, 2008
Human Reproduction, doi:10.1093/humrep/den280
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© The Author 2008. Published by Oxford University Press on behalf of the European Society of Human Reproduction and Embryology. All rights reserved. For Permissions, please email: journals.permissions@oxfordjournals.org

The pattern of localization of the putative oocyte activation factor, phospholipase C {zeta}, in uncapacitated, capacitated, and ionophore-treated human spermatozoa

P. Grasa1,{dagger}, K. Coward1,2,{dagger}, C. Young1,{dagger} and J. Parrington1,3

1 Department of Pharmacology, University of Oxford, Mansfield Road, Oxford OX1 3QT, UK 2 Present address: Nuffield Department of Obstetrics and Gynaecology, University of Oxford, Level 3, Women's Centre, John Radcliffe Hospital, Headington, Oxford OX3 9DU, UK

3 Correspondence address. E-mail: john.parrington{at}pharm.ox.ac.uk

BACKGROUND: Recent studies suggest that in mammals, oocyte activation at fertilization is triggered by a sperm-specific phospholipase C, PLC{zeta}. We investigated PLC{zeta} localization in human spermatozoa.

METHODS: A polyclonal antibody was generated against human PLC{zeta} and used in immunoblotting and immunofluorescence studies of ejaculated human sperm in uncapacitated and capacitated states. An ionophore was also used to induce the acrosome reaction in vitro.

RESULTS: After verifying specificity of the anti-PLC{zeta} antibody by immunoblotting, immunofluorescence studies showed that the predominant localization of PLC{zeta} in uncapacitated sperm was in the equatorial region, a pattern maintained following capacitation and ionophore treatment. The analysis of pooled samples showed ~88% of uncapacitated sperm expressed PLC{zeta} in the equatorial region, whereas ~35% and ~21% of sperm expressed additional populations of PLC{zeta} in the acrosomal or post-acrosomal region, respectively. One population of PLC{zeta} was observed in the post-acrosomal region of ~12% of sperm. The proportion of cells with post-acrosomal PLC{zeta} increased following capacitation and ionophore treatment (P < 0.05). The same tendency was found in individual samples. There was a strong correlation (r = 0.716, P < 0.0001) between presence of an intact acrosome and proportion of sperm immunoreactive to PLC{zeta} in the acrosomal region.

CONCLUSIONS: PLC{zeta} was variably detectable in three localities within the sperm head: the equatorial segment and acrosomal/post-acrosomal region. Variability in PLC{zeta} localization in sperm from fertile males may reflect differences in oocyte activation capabilities between individuals or within an ejaculate. This approach may help in investigating the possible links between PLC{zeta} and certain types of male infertility.

Key words: oocyte activation/phospholipase C zeta/sperm/capacitation/acrosome reaction

{dagger} These authors contributed equally.

Submitted on March 7, 2008; resubmitted on June 18, 2008; accepted on June 24, 2008.


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