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BOR - Papers in Press, published online ahead of print November 27, 2002.
Biol Reprod 2002, 10.1095/biolreprod.102.011023
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biolreprod.102.011023v1
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BIOLOGY OF REPRODUCTION 68, 1463–1469 (2003)
DOI: 10.1095/biolreprod.102.011023
© 2003 by the Society for the Study of Reproduction, Inc.

Gamete Biology

Localization of Tyrosine Phosphorylated Proteins in Human Sperm and Relation to Capacitation and Zona Pellucida Binding1

D. Sakkasa, G. Leppens-Luisierb, H. Lucasb, D. Chardonnensb, A. Campanab, D.R. Frankenc, and F. Urner2,b

a Department of Obstetrics and Gynecology, Yale University School of Medicine, New Haven, Connecticut 06520 b Clinic of Sterility, Department of Obstetrics and Gynecology, University Hospital of Geneva, 1211 Geneva 14, Switzerland c Department of Obstetrics and Gynecology, Reproductive Biology Unit, University of Stellenbosch and Tygerberg Hospital, Tygerberg, South Africa

Mammalian sperm must undergo a process known as capacitation before fertilization can take place. A key intracellular event that occurs during capacitation is protein tyrosine phosphorylation. The objective of this study was to investigate and visualize protein tyrosine phosphorylation patterns in human sperm during capacitation and interaction with the zona pellucida. The presence of specific patterns was also assessed in relation to the fertilizing capacity of the spermatozoa after in vitro fertilization. Protein tyrosine phosphorylation was investigated by immunofluorescence. Phosphorylation increased significantly with capacitation and was localized mainly to the principal piece of human sperm. Following binding to the zona pellucida, the percentage of sperm with phosphotyrosine residues localized to both the neck and the principal piece was significantly higher in bound sperm than in capacitated sperm in suspension. When the percentage of principal piece-positive sperm present after capacitation was <7%, fertilization rates after in vitro fertilization were reduced. Different compartments of human spermatozoa undergo a specific sequence of phosphorylation during both capacitation and upon binding to the zona pellucida. Tyrosine phosphorylation in the principal and neck piece may be considered a prerequisite for fertilization in humans.

1 This work was supported by the Fonds National Suisse (32-55693.98).

2 Correspondence: Françoise Urner, Laboratoire des Gamètes, Clinique de Stérilité, Hôpital Cantonal, 30, Bd de la Cluse, 1211 Geneva 14, Switzerland. FAX: 41 22 38 24 385; francoise.urner{at}hcuge.ch




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