Biochemical Studies of the Envelope Transformations in XenopusLaevis Eggs

  • George L. Gerton


The envelopes that enclose the eggs of the amphibian Xenopus laevis were isolated and examined for biochemical correlates of the ultrastructural and sperm penetrability differences among the coelomic egg envelope (CE), the vitelline envelope (VE), and the fertilization envelope (FE). By sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), the 43,000 molecular weight glycoproteins of CEs were found to be converted to components with molecular weights of 41,000 in VEs; also, a protein with a molecular weight of 57,000 was added to the envelope during the CE-to-VE conversion. The molecular weights of two components decreased during the VE-to-FE conversion, from 69,000 and 64,000 in the VE to 66,000 and 61,000 in the FE. Components from the cortical granules and the innermost jelly coat were also added to the newly formed FE. As detected by iodination with lactoperoxidase or IODOGEN, both the CE-to-VE and the VE-to-FE conversions caused conformational changes in envelope glycoproteins. Peptide mapping demonstrated that the 43,000 molecular weight components of CE were precursors to the 41,000 molecular weight components of VE and the 69,000 and 64,000 molecular weight components of VE were precursors to the 66,000 and 61,000 molecular weight components of FE. The CE-to-VE conversion presumably occurs in the first portion of the oviduct. Experiments probing the VE-to-FE conversion demonstrated the need for an intact jelly coat for the molecular weight changes to occur. Sperm were not required for the envelope alteration; the SDS-PAGE pattern of envelopes from jellied eggs activated with the Ca++-ionophore A23187 were indistinguishable from the FE. These studies show that there are molecular correlates of the morphological and biological differences among the envelopes. The CE-to-VE and the VE-to-FE conversions follow a similar pattern: in both cases, material is added to the envelope and there are changes in the molecular weights of some of the components.


Xenopus Laevis Acrosome Reaction Cortical Granule Perivitelline Space Jelly Coat 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Birr, C.A. 1979. Immunoelectophoretic studies of the jelly coat ligand for the cortical granule lectin of Xenopus laevis eggs. Ph.D. Thesis, University of California, Davis.Google Scholar
  2. 2.
    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.CrossRefGoogle Scholar
  3. 3.
    Carroll, E.J., Jr., and Epel, D. 1975. Isolation and biological activity of the proteases released by sea urchin eggs following fertilization. Develop. Biol. 44: 22–32.CrossRefGoogle Scholar
  4. 4.
    Carroll, E.J., Jr., and Hedrick, J.L. 1974. Hatching in the toad Xenopus laevis: Morphological events and evidence for a hatching enzyme. Develop. Biol. 38: 1–13.CrossRefGoogle Scholar
  5. 5.
    Cleveland, D.W., Fischer, S.G., Kirschner, M.W., and Laemmli, U.K. 1977. Peptide mapping by limited proteolysis in sodium dodecyl sulfate and analysis by gel electrophoresis. J. Biol. Chem. 252: 1102–1106.Google Scholar
  6. 6.
    Eckhardt, A.E., Hayes, C.E., and Goldstein, I.J. 1976. A sensitive method for the detection of glycoproteins in polyacrylamide gels. Anal. Biochem. 73: 192–197.CrossRefGoogle Scholar
  7. 7.
    Elinson, R.P. 1973. Fertilization of frog body cavity eggs enhanced by treatments affecting the vitelline coat. J. Exp. Zool. 183: 291–302.CrossRefGoogle Scholar
  8. 8.
    Florman, H., and Storey, B. 1982. Mouse gamete interactions: The zona pellucida is the site of the acrosome reaction leading to fertilization in vitro. Develop. Biol. 91: 121–130.CrossRefGoogle Scholar
  9. 9.
    Foerder, C. A., and Shapiro, B.M. 1977. Release of ovoperoxidase from sea urchin eggs hardens the fertilization membrane with tyrosine crosslinks. Proc. Natl. Acad. Sci., USA 74: 4214–4218.ADSCrossRefGoogle Scholar
  10. 10.
    Gerton, G.L., Wardrip, N.J., Hedrick, J.L. 1982. A gel eluter for the recovery of proteins separated by polyacrylamide gel electrophoresis. Anal. Biochem. 126: 116–121.CrossRefGoogle Scholar
  11. 10.
    Gerton, G.L., Wardrip, N.J., Hedrick, J.L. 1982. A gel eluter for the recovery of proteins separated by polyacrylamide gel electrophoresis. Anal. Biochem. 126: 116–121.CrossRefGoogle Scholar
  12. 10.
    Gerton, G.L., Wardrip, N.J., Hedrick, J.L. 1982. A gel eluter for the recovery of proteins separated by polyacrylamide gel electrophoresis. Anal. Biochem. 126: 116–121.CrossRefGoogle Scholar
  13. 13.
    Glabe, C.G., and Vacquier, V.D. 1978. Egg surface glycoprotein receptor for sea urchin sperm binding. Proc. Natl. Acad. Sci., USA 75: 881–885.ADSCrossRefGoogle Scholar
  14. 14.
    Greve, L.C., and Hedrick, J.L. 1978. An immunocytochemical localization of the cortical granule lectin in fertilized and unfertilized eggs of Xenopus laevis. Gamete Res. l:13–18.Google Scholar
  15. 15.
    Grey, R.D., Wolf, D.P., and Hedrick, J.L. 1974. Formation and structure of the fertilization envelope in Xenopus laevis. Develop. Biol. 36: 44–61.CrossRefGoogle Scholar
  16. 16.
    Grey, R.D., Working, P.K., and Hedrick, J.L. 1976. Evidence that the fertilization envelope blocks sperm entry in eggs of Xenopus laevis: Interactiqn of sperm with isolated envelopes. Develop. Biol. 54: 52–60.CrossRefGoogle Scholar
  17. 17.
    Grey, R.D., Working, P.K., and Hedrick, J.L. 1977. Alteration of structure and penetrability of the vitelline envelope after passage of eggs from coelom tp oviduct in Xenopus laevis. J. Exp. Zool. 201: 73–83.CrossRefGoogle Scholar
  18. 18.
    Grey, R.D., Bastiani, M.J., Webb, D.J., and Schertel, E.R. 1982. An electrical block is required to prevent polyspermy in eggs fertilized by natural mating of Xenopus laevis. Develop. Biol. 89: 475–484.Google Scholar
  19. 19.
    Gwatkin, R.B.L., and Williams, D.T. 1977. Receptor activity of the hamster and mouse solubilized zona pellucida before and after the zona reaction. J. Reprod. Fert. 49: 55–59.CrossRefGoogle Scholar
  20. 20.
    Hedrick, J.L., Smith, A.J., Yurewicz, E.C., Oliphant. G., and Wolf, D.P. 1974. The incorporation and fate of [35S]-sulfate in the jelly coat of Xenopus laevis eggs. Biol. Repro. 11: 534–542.CrossRefGoogle Scholar
  21. 21.
    Katagiri, C. 1974. A high frequency of fertilization in premature and mature coelomic toad eggs after enzymic renjovl of vitelline membrane. J. Embryol. Exp. Morphol. 31:573–587.Google Scholar
  22. 22.
    Katagiri, C., Iwaot Y., and Yoshizaki, N. 1982. Participation of oviducal pars recta secretions in inducing the acrosome reaction and release of vitelline coat lysin in fertilizing toad sperm. Develop. Biol. 94: 1–10.CrossRefGoogle Scholar
  23. 23.
    Kutryk, M.A., and Hedrick, J.L. 1983. Melting of glycoprotein egg envelopes as a probe of supramolecular conformational differences. Fed. Proc. 42: 1996.Google Scholar
  24. 24.
    Landon, M. 1977. Cleavage at aspartyl-prolyl peptide bonds. Methods Enzymol. 47: 145–149.CrossRefGoogle Scholar
  25. 25.
    Miceli, D.C., Fernandez, S.N., Raisman, J.S., and Barbieri, F.D. 1978a. A trypsin-like oviducal proteinase involved in Bufo arenarum fertilization. J. Embryol. Exp. Morphol. 48: 79–91.Google Scholar
  26. 26.
    Miceli, D.C., Fernandez, S.N., and del Pino, E.J. 1978b. An oviducal enzyme isolated by affinity chromatography which acts upon the vitelline envelope of Bufo arenarum oocytes. Biochim. Biophys. Acta. 526: 289–292.Google Scholar
  27. 27.
    Miceli, D.C., and Fernandez, S.N. 1982. Properties of an oviducal protein involved in amphibian oocyte fertilization. J. Exp. Zool. 221: 357–364.CrossRefGoogle Scholar
  28. 28.
    Nishihara, T., and Hedrick, J.L. 1977a. A molecular mechanism for envelope elevation at fertilization. Fed. Proc. 36: 811.Google Scholar
  29. 29.
    Nishihara, T., and Hedrick, J.L. 1977b. Reconstruction of the fertilization envelope from its component parts. J. Cell Biol. 75: 172a.Google Scholar
  30. 30.
    Nishihara, T., Gerton, G.L., and Hedrick, J.L. 1983. Radioiodination studies of the envelopes from Xenopus laevis eggs. J. Cell. Biochem. 22: 235–244.CrossRefGoogle Scholar
  31. 31.
    SeGall, G.K., and Lennarz, W.J. 1979. Chemical characterization of the component of the jelly coat from sea urchin eggs responsible for the induction of the acrosome reaction. Develop. Biol. 71: 33–48.CrossRefGoogle Scholar
  32. 32.
    Vacquier, V.D., Tegner, M.J., and Epel, D. 1973. Protease released from sea urchin eggs at fertilization alters the vitelline layer and aids in preventing polyspermy. Exp. Cell Res. 80: 111–119.CrossRefGoogle Scholar
  33. 33.
    Weiner, A.M., Piatt, T., and Weber, K. 1972. Amino-terminal sequence analysis of proteins purified on a nanomole scale by gel electrophoresis. J. Biol. Chem. 247: 3242–3251.Google Scholar
  34. 34.
    Wolf, D.P. 1974. On the contents of the cortical granules from Xenopus laevis eggs. Develop. Biol. 38: 14–29.CrossRefGoogle Scholar
  35. 35.
    Wolf, D.P., and Hedrick, J.L. 1971a. A molecular approach to fertilization. II. Viability and artificial fertilization of Xenopus laevis gametes. Develop. Biol. 25: 348–359.CrossRefGoogle Scholar
  36. 36.
    Wolf, D.P., and Hedrick, J.L. 1971b. A molecular approach to fertilization. III. Development of a bioassay for sperm capacitation. Develop. Biol. 25: 360–376.CrossRefGoogle Scholar
  37. 37.
    Wolf, D.P., Nishihara, T., West, D.M., and Hedrick, J.L. 1976. Isolation, physicochemical properties, and macromolecular composition of the vitelline and fertilization envelopes from Xenopus laevis eggs. Biochemistry 15: 3671–3677.CrossRefGoogle Scholar
  38. 38.
    Wyrick, R.E., Nishihara, T., and Hedrick, J.L. 1974. Agglutination of jelly coat and cortical granule components and the block to polyspermy in the amphibian Xenopus laevis. Proc. Natl. Acad. Sci., USA 71: 2067–2071.ADSCrossRefGoogle Scholar
  39. 39.
    Yurewicz, E.C., Oliphant, G.,and Hedrick, J.L. 1975. The Xenopus laevis egg jelly coat. macromolecular composition Biochemistry 14: 3101–3107.Google Scholar

Copyright information

© Plenum Press, New York 1986

Authors and Affiliations

  • George L. Gerton
    • 1
  1. 1.Division of Reproductive Biology, Department of Obstetrics and GynecologyUniversity of Pennsylvania School of MedicinePhiladelphiaUSA

Personalised recommendations