Molecular Dissection of the Mouse Zona Pellucida

An Electron-Microscopic Perspective Utilizing High-Resolution Colloidal-Gold Labeling Methods
  • Frederick W. K. Kan
Part of the Methods in Molecular Biology book series (MIMB, volume 347)


The zona pellucida (ZP) is an extracellular coat that encloses growing oocytes, ovulated eggs, and preimplantation embryos in mammals. The ZP contains receptors that mediate initial interactions between the sperm and egg and the relative species-specificity during gamete interaction. It also prevents polyspermy and protects the developing embryo prior to implantation. The current model of the organization of mouse ZP depicts this extracellular matrix as an extensive three-dimensional array of long interconnected filaments of a structural repeat. Using high-resolution colloidal-gold labeling methods in combination with specific lectins and monoclonal antibodies against three major ZP glycoproteins, we have characterized the outer and inner mouse ZP during folliculogenesis and shown the modifications of the ZP after ovulation and fertilization. Our immunogold labeling results also indicate the involvement of a vesicular aggregate, a specialized subcellular compartment in the oocyte, in the synthesis and secretion of ZP glycoproteins.


  1. 1.
    Shimizu, S., Tsuji, M., and Dean, J. (1983) In vitro biosynthesis of three sulfated glycoproteins of murine zonae pellucidae by oocytes grown in follicle culture. J. Biol. Chem. 258, 5858–5863.PubMedGoogle Scholar
  2. 2.
    Wassarman, P. M. (1988) Zona pellucida glycoproteins. Annu. Rev. Biochem. 57,415–442.CrossRefPubMedGoogle Scholar
  3. 3.
    Dunbar, B. S., Avery, S., Lee, V., et al. (1994) The mammalian zona pellucida: its biochemistry, immunochemistry, molecular biology and developmental expression. Reprod. Fertil. Dev. 6, 59–76.CrossRefGoogle Scholar
  4. 4.
    Bleil, J. D. and Wassarman, P. M. (1980) Mammalian sperm-egg interaction: identification of a glycoprotein in mouse egg zonae pellucidae possessing receptor activity for sperm. Cell 20, 873–882.CrossRefPubMedGoogle Scholar
  5. 5.
    Bleil, J. D. and Wassarman, P. M. (1988) Galactose at the nonreducing terminus of O-linked oligosaccharides of mouse egg zona glycoprotein ZP3 is essential for the glycoprotein’s sperm receptor activity. Proc. Natl. Acad. Sci. USA 85, 6778–6782.CrossRefPubMedGoogle Scholar
  6. 6.
    Bleil, J. D., Greve, J. M., and Wassarman, P. M. (1988) Identification of a secondary sperm receptor in the mouse egg zona pellucida: role in maintenance of binding of acrosome-reacted sperm to eggs. Dev. Biol. 128, 376–385.CrossRefPubMedGoogle Scholar
  7. 7.
    Greve, J. M. and Wassarman, P. M. (1988) Mouse egg extracellular coat is a matrix of interconnected filaments possessing a structural repeat. J. Mol. Biol. 181, 253–264.CrossRefGoogle Scholar
  8. 8.
    Rankin, T. and Dean, J. (2000) The zona pellucida: using molecular genetics to study the mammalian egg coat. Rev. Reprod. 5, 114–121.CrossRefPubMedGoogle Scholar
  9. 9.
    Hoodbhoy, T. and Dean, J. (2004) Insights into the molecular basis of sperm-egg recognition in mammals. Reproduction 127, 417–422.CrossRefPubMedGoogle Scholar
  10. 10.
    East, I. J. and Dean, J. (1984) Monoclonal antibodies as probes of the distribution of ZP-2, the major sulfated glycoprotein of the murine zona pellucida. J. Cell Biol. 98, 795–800.CrossRefPubMedGoogle Scholar
  11. 11.
    East, J. D., Gulyas, B. J., and Dean, J. (1985) Monoclonal antibodies to murine zona pellucida protein with sperm receptor activity: effects on fertilization and early development. Dev. Biol. 109, 268–273.CrossRefPubMedGoogle Scholar
  12. 12.
    Rankin, T. L., Tong, Z.-B., Castle, P. E., et al. (1998) Human ZP3 restores fertility in Zp3 null mice without affecting order-specific sperm binding. Development 125, 2415–2424.PubMedGoogle Scholar
  13. 13.
    El-Mestrah, M., Castle, P. E., Borossa, G., and Kan, F. W. K. (2002) Subcellular distribution of ZP1, ZP2, and ZP3 glycoproteins during folliculogenesis and demonstration of their topographical disposition within the zona matrix of mouse ovarian oocytes. Biol. Reprod. 66, 866–876.CrossRefPubMedGoogle Scholar
  14. 14.
    El-Mestrah, M. and Kan, F. W. K. (2001) Distribution of lectin-binding glycosidic residues in the hamster follicular oocytes and their modifications in the zona pellucida after ovulaton. Mol. Reprod. Dev. 60, 517–534.CrossRefPubMedGoogle Scholar
  15. 15.
    El-Mestrah, M. and Kan, F. W. K. (2002) Variation in modifications of sugar residues in hamster zona pellucida after in vivo fertilization and in vitro egg activation. Reproduction 123, 671–682.CrossRefPubMedGoogle Scholar
  16. 16.
    Avilé, M., El-Mestrah, M., Jaber, L., Castells, M. T., Ballesta, J., and Kan, F. W. K. (2000) Cytochemical demonstration of modification of carbohydrates in the mouse zona pellucida during folliculogenesis. Histochem. Cell Biol. 113, 207–219.CrossRefGoogle Scholar
  17. 17.
    Frens, G. (1973) Controlled nucleation for the regulation of the particle size in monodispersed gold suspension. Nature Phys. Sci. 241, 20–22.Google Scholar
  18. 18.
    Bendayan, M., Nanci, A.,and Kan, F. W. K. (1987) Effect of tissue processing on colloidal gold cytochemistry. J. Histochem. Cytochem. 35, 983–996.PubMedGoogle Scholar

Copyright information

© Humana Press Inc. 2006

Authors and Affiliations

  • Frederick W. K. Kan
    • 1
  1. 1.Department of Anatomy and Cell BiologyQueen’s UniversityKingstonCanada

Personalised recommendations