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Biological Effects of Recombinant Human Zona Pellucida Proteins on Sperm Function¹

Center for Research on Reproduction and Women's Health,⁴ University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania 19104-6080 Departamento de Biología de la Reproducción,⁵ Instituto Nacional de Ciencias Médicas y Nutrición Salvador Zubirán, Mexico City 14000, Mexico

ABSTRACT

The initial interaction between gametes takes place at the level of the sperm surface and the zona pellucida (ZP), the extracellular matrix of the egg in mammals. Successful fertilization requires the proper molecular recognition of the ZP by the sperm. Recently, human ZP was demonstrated to be composed of four proteins: ZP1, ZP2, ZP3, and ZP4. The goals of this study were to determine the effects of recombinant human ZP2, ZP3, and ZP4 on human sperm acrosomal exocytosis and sperm motility. Exposure of sperm to ZP proteins, alone or in combination, promoted acrosomal exocytosis in a time-dependent manner. This effect occurred in parallel with a considerable decrease in progressive motility, coincident with an increase in nonprogressive sperm motility. An analysis of kinetic parameters of ZP-treated sperm demonstrated that a characteristic motility pattern could be defined by values of curvilinear velocity > 63.9 µm/s and linearity 15.5%. A strong correlation between curvilinear velocity and the amplitude of lateral head displacement was also observed. The incidence of sperm having these particular kinetic parameters increased after exposure to ZP proteins. These studies of two processes involved in sperm penetration through the ZP confirm that zona glycoproteins promote acrosomal exocytosis and now establish an additional role for these components as modifiers of sperm motility.

acrosome reaction, acrosomal exocytosis, fertilization, gamete biology, sperm capacitation, sperm motility and transport, zona pellucida

¹ Supported by Contraceptive Research and Development (CONRAD) Twinning Program grants MFG-02–62 and MFG-02–63, NIH HD-41552, and the National Council of Science and Technology (CONACyT, Mexico). Also supported in part by the Bill and Melinda Gates Foundation (to X.J.F.). P.C.C. is a recipient of a training fellowship from Fundación Salud 2000 and Fundación Tambre, Madrid, Spain. M.E.G.G. and M.G.C. have been supported by CONACyT, Mexico.

² Correspondence: George L. Gerton, Center for Research on Reproduction and Women's Health, Department of Obstetrics and Gynecology, University of Pennsylvania Medical Center, 421 Curie Blvd., 1311 BRB II/III, Philadelphia, PA 19104-6080. FAX: 215 573 7627; gerton{at}mail.med.upenn.edu

³ These authors contributed equally to this work.