Abstract
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.
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References
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.
Wassarman, P. M. (1988) Zona pellucida glycoproteins. Annu. Rev. Biochem. 57,415–442.
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.
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.
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.
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.
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.
Rankin, T. and Dean, J. (2000) The zona pellucida: using molecular genetics to study the mammalian egg coat. Rev. Reprod. 5, 114–121.
Hoodbhoy, T. and Dean, J. (2004) Insights into the molecular basis of sperm-egg recognition in mammals. Reproduction 127, 417–422.
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.
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.
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.
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.
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.
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.
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.
Frens, G. (1973) Controlled nucleation for the regulation of the particle size in monodispersed gold suspension. Nature Phys. Sci. 241, 20–22.
Bendayan, M., Nanci, A.,and Kan, F. W. K. (1987) Effect of tissue processing on colloidal gold cytochemistry. J. Histochem. Cytochem. 35, 983–996.
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Kan, F.W.K. (2006). Molecular Dissection of the Mouse Zona Pellucida. In: Brockhausen, I. (eds) Glycobiology Protocols. Methods in Molecular Biology, vol 347. Humana Press. https://doi.org/10.1385/1-59745-167-3:289
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DOI: https://doi.org/10.1385/1-59745-167-3:289
Publisher Name: Humana Press
Print ISBN: 978-1-58829-553-8
Online ISBN: 978-1-59745-167-3
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