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Mammalian Fertilization Is Dependent on Multiple Membrane Fusion Events*

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Cell Fusion

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 475))

Summary

Successful completion of fertilization in mammals is dependent on three membrane fusion events. These are (1) the acrosome reaction of sperm, (2) the fusion of sperm and egg plasma membranes to form a zygote, and (3) the cortical reaction of fertilized eggs. Extensive research into the molecular basis of each of these events has identified candidate proteins and factors involved in fusion of membranes during the mammalian fertilization process. Some of this information is provided here.

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References

  1. Alberts, B., Johnson, A., Lewis, J., Raff, M., Roberts, K., and Walter, P. (2002) Molecular Biology of the Cell (4th ed.). Garland Science, New York.

    Google Scholar 

  2. Jahn, R., Lang, T., and Sudhof, T. C. (2003) Membrane fusion. Cell 112, 519–533.

    Article  CAS  PubMed  Google Scholar 

  3. Lodish, H., Berk, A., Matsudaira, P., Kaiser, C. A., Krieger, M., Scott, M. P., Zipursky, S. L., and Darnell, J. (2004) Molecular Cell Biology (5th ed.). Freeman, New York.

    Google Scholar 

  4. Evans, J. P. and Florman, H. M. (2002) The state of the union: the cell biology of fertilization. Nat. Cell Biol. 4 (Suppl. 1), S57–S63.

    Article  PubMed  Google Scholar 

  5. Evans, J. P. (2002) The molecular basis of sperm–oocyte membrane interactions during mammalian fertilization. Hum. Reprod. Update 8, 297–311.

    Article  CAS  PubMed  Google Scholar 

  6. Florman, H. M. and Ducibella, T. (2006) Fertilization in mammals, in The Physiology of Reproduction (Neill J.D., ed.), vol. 1. Elsevier, San Diego, pp. 55–112.

    Google Scholar 

  7. Primakoff, P. and Myles, D. G. (2002) Gamete fusion in mammals, in Fertilization (Hardy, D. M., ed.). Academic Press, San Diego, pp. 303–318.

    Chapter  Google Scholar 

  8. Ramalho-Santos, J., Schatten, G., and Moreno, R. D. (2002) Control of membrane fusion during spermiogenesis and the acrosome reaction. Biol. Reprod. 69, 254–260.

    Google Scholar 

  9. Stein, K. K., Primakoff, P., and Myles, D. (2004) Sperm–egg fusion: events at the plasma membrane. J. Cell Sci. 117, 6269–6274.

    Article  CAS  PubMed  Google Scholar 

  10. Wassarman, P. M. (1999) Mammalian fertilization: molecular aspects of gamete adhesion, exocytosis, and fusion. Cell 96, 175–183.

    Article  CAS  PubMed  Google Scholar 

  11. Wassarman, P. M., Jovine, L., and Litscher, E. S. (2001) A profile of fertilization in mammals. Nat. Cell Biol. 3, E59–E64.

    Article  CAS  PubMed  Google Scholar 

  12. Chernomordik, L. V. and Kozlov, M. M. (2003) Protein–lipid interplay in fusion and fission of biological membranes. Annu. Rev. Biochem. 72, 175–207.

    Article  CAS  PubMed  Google Scholar 

  13. Jahn, R. and Grubmuller, H. (2002) Membrane fusion. Curr. Opin. Cell Biol. 14, 488–495.

    Article  CAS  PubMed  Google Scholar 

  14. Mayer, A. (2002) Membrane fusion in eukaryotic cells. Annu. Rev. Cell Dev. Biol. 18, 289–314.

    Article  CAS  PubMed  Google Scholar 

  15. Eddy, E. M. (2006) The spermatozoon, in The Physiology of Reproduction (Neill J. D., ed.), vol. 1. Elsevier, San Diego, pp. 3–54.

    Google Scholar 

  16. Gerton, G. L. (2002) Function of the sperm acrosome, in Fertilization (Hardy, D. M., ed.). Academic Press, San Diego, pp. 265–302.

    Chapter  Google Scholar 

  17. Yanagimachi, R. (1994) Mammalian fertilization, in The Physiology of Reproduction (Knobil E. and Neill J. D., eds.), vol. 1. Raven Press, New York, pp. 189–317.

    Google Scholar 

  18. Wassarman, P. M. and Albertini, D. F. (1994) Cellular and molecular biology of the mammalian ovum, in The Physiology of Reproduction (Knobil, E. Neill, J. D., eds.), vol. 1. Raven Press, N Y, pp. 79–122.

    Google Scholar 

  19. Yanagimachi, R. (1981) Mechanisms of fertilization in mammals, in Fertilization and Embryonic Development In Vitro (Mastroianni L. and Biggers J. D., eds.). Plenum Press, New York, pp. 81–187.

    Google Scholar 

  20. Jaiswal, B. S. and Eisenbach, M. (2002) Capacitation, in Fertilization (Hardy, D. M., ed.). Academic Press, San Diego, pp. 57–117.

    Chapter  Google Scholar 

  21. Kim, K.-S. and Gerton, G. L. (2003) Differential release of soluble and matrix components: evidence for intermediate states of secretion during spontaneous acro- somal exocytosis in mouse sperm. Dev. Biol. 264, 141–152.

    Article  CAS  PubMed  Google Scholar 

  22. Kopf, G. S., Ning, X. P., Visconti, P. E., Purdon, M., Galantino-Homer, H., and Fornes, M. (1999) Signaling mechanisms controlling mammalian sperm fertiliza- tion competence and activation. In The Male Gamete (Gagnon C., ed.) 105–118. Cache River Press, Vienna, IL.

    Google Scholar 

  23. Florman, H. M., Kirkman-Brown, J., Brown, J. C., Jungnickel, M. K., and Sutton, K. A. (2003) The acrosome reaction: an example of egg-activated signal transduc- tion in sperm. ChemTracts Biochem. Mol. Biol. 16, 126–133.

    CAS  Google Scholar 

  24. Kopf, G. S. (2002) Signal transduction mechanisms regulating sperm acrosomal exocytosis, in Fertilization (Hardy, D. M., ed.). Academic Press, San Diego, pp. 181–223.

    Chapter  Google Scholar 

  25. Wassarman, P. M., Bleil, J. D., Florman, H. M., Greve, J. M., Roller, R. J., Salzmann, G. S., and Samuels, F. G. (1985) The mouse egg 's sperm receptor: what is it and how does it work? Cold Spring Harb. Symp. Quant. Biol. 50, 11–19.

    CAS  PubMed  Google Scholar 

  26. Arnoult, C., Zeng, Y. , and Florman, H. M. (1996) ZP3-dependent activation of sperm cation channels regulates acrosomal secretion during mammalian fertilization. J. Cell Biol. 134, 637–645.

    Article  CAS  PubMed  Google Scholar 

  27. Darszon, A., Acevedo, J. J., Galindo, B. E., Hernandez-Gonzalez, E. O., Nishigaki, T., Trevino, C. L., Wood, C., and Beltran, C. (2006) Sperm channel diversity and functional multiplicity. Reproduction 131, 977–988.

    Article  CAS  PubMed  Google Scholar 

  28. Jungnickel, M. K., Marrero, H., Birnbaumer, L., Lemos, J. R., and Florman, H. M. (2001) Trp2 regulates entry of Ca2+ into mouse sperm triggered by egg ZP3. Nat. Cell Biol. 5, 499–502.

    Article  Google Scholar 

  29. De Blas, G. A., Roggero, C. M., Tomes, C. N., and Mayorga, L. S. (2005) Dynamics of SNARE assembly and disassembly during sperm acrosomal exocytosis. PloS Biol. 3, e323.

    Article  PubMed  Google Scholar 

  30. Iida, H., Yoshinaga, Y., Tanaka, S., Toshimori, K., and Mori, T. (1999) Identification of Rab3A GTPase as an acrosome-associated small GTP-binding protein in rat sperm. Dev. Biol. 211, 144–155.

    Article  CAS  PubMed  Google Scholar 

  31. Ramalho-Santos, J., Moreno, R. D., Sutovsky, P., Chan, A. W., Hewitson, L., Wessel, G. M., Simerly, C. R., and Schatten, G. (2000) SNAREs in mammalian sperm: possible implications for fertilization. Dev. Biol. 223, 54–69.

    Article  CAS  PubMed  Google Scholar 

  32. Tomes, C. N., Michaut, M., De Blas, G. A., Visconti, P., Matti, U., and Mayorga, L. S. (2002) SNARE complex assembly is required for human sperm acrosome reaction. Dev. Biol. 243, 326–338.

    Article  CAS  PubMed  Google Scholar 

  33. Tomes, C. N., De Blas, G. A., Michaut, M. A., Farre, E. V., Cherhitin, O., Visconti, P. E., and Mayorga, L. S. (2005) Alpha-SNAP and NSF are required in a priming step during the human sperm acrosome reaction. Mol. Hum. Reprod. 11, 43–51.

    Article  CAS  PubMed  Google Scholar 

  34. Yunes, R., Michaut, M., Tomes, C., and Mayorga, L. S. (2000) Rab3A triggers the acrosome reaction in permeabilized human spermatozoa. Biol. Reprod. 62, 1084–1089.

    Article  CAS  PubMed  Google Scholar 

  35. Howes, E. A. and Jones, R. (2003) Secondary binding of mammalian sperm to the zona pellucida. ChemTracts Biochem. Mol. Biol. 16, 134–141.

    CAS  Google Scholar 

  36. Oh, E., Wortzman, G. B., Zhu, X., and Evans, J. P. (2003) Getting sperm and egg together: the molecules of gamete membrane interactions. ChemTracts Biochem. Mol. Biol. 16, 142–157.

    CAS  Google Scholar 

  37. White, J. M. (2003) ADAMs: modulators of cell–cell and cell–matrix interactions. Curr. Opin. Cell Biol. 15, 598–606.

    Article  CAS  PubMed  Google Scholar 

  38. Cho, C., Bunch, D. O., Faure, J.-E., Goulding, E. H., Eddy, E. M., Primakoff, P., and Myles, D. G. (1998) Fertilization defects in sperm from mice lacking fertilin β. Science 281, 1857–1859.

    Article  CAS  Google Scholar 

  39. Cho, C., Ge H., Branciforte, D., Primakoff, P., and Myles, D. G. (2000) Analysis of mouse fertilin in wild-type and fertilin β−/− sperm: evidence for C-terminal modification, α / β dimerization, and lack of essential role of fertilin b in sperm–egg fusion. Dev. Biol. 222, 289–295.

    Article  CAS  PubMed  Google Scholar 

  40. He, Z. Y., Brakefusch, C., Fassler, R., Kreidberg, J. A., Primakoff, P. , and Myles, D. G. (2003) None of the integrins known to be present on the mouse egg or to be ADAM receptors are essential for sperm–egg binding and fusion. Dev. Biol. 254, 226–237.

    Article  CAS  PubMed  Google Scholar 

  41. Kim, E., Yamashita, M., Nakanishi, T., Park, K.-E., Kimura, M., Kashiwabara, S.-I., and Baba, T. (2006) Mouse sperm lacking ADAM1β /ADAM2 fertilin can fuse with the egg plasma membrane and effect fertilization. J. Biol. Chem. 281, 5634–5639.

    Article  CAS  PubMed  Google Scholar 

  42. Miller, B. J., Georges-Labouesse, E., Primakoff, P., and Myles, D. G. (2000) Normal fertilization occurs with eggs lacking the integrin α 6 β1 and is CD9-depen-dent. J. Cell Biol. 149, 1289–1295.

    Article  CAS  PubMed  Google Scholar 

  43. Nishimura, H., Cho, C., Branciforte, D. R., Myles, D. G., and Primakoff, P. (2001) Analysis of loss of adhesive function in sperm lacking cyritestin or fertilin β. Dev. Biol. 233, 204–213.

    Article  CAS  Google Scholar 

  44. Shamsadin, R., Adham, I.M., Nayernia, K., Heinlein, U. A. O., Oberwinkler, H., and Engel, W. (1999) Male mice deficient for germ-cell cyritestin are infertile. Biol. Reprod. 61, 1445–1451.

    Article  CAS  PubMed  Google Scholar 

  45. Stein, K. K., Go, J. C., Lane, W. S., Primakoff, P., and Myles, D. G. (2006) Proteomic analysis of sperm regions that mediate sperm–egg interactions. Proteomics 6, 3533–3543.

    Article  CAS  PubMed  Google Scholar 

  46. Hemler, M. E. (1998) Integrin associated proteins. Curr. Opin. Cell Biol. 10, 578–585.

    Article  CAS  PubMed  Google Scholar 

  47. Porter, J. C. and Hogg, N. (1998) Integrins take partners: cross-talk between integ- rins and other membrane receptors. Trends Cell Biol. 8, 390–396.

    Article  CAS  PubMed  Google Scholar 

  48. Tachibana, I. and Hemler, M. E. (1999) Role of transmembrane 4 superfamily (TM4SF) proteins CD9 and CD81 in muscle cell fusion and myotube maintenance. J. Cell Biol. 146, 893–904.

    Article  CAS  PubMed  Google Scholar 

  49. Kaji, K., Odu, S., Shikano, T., Ohnuki, T., Uematsu, Y. , Sakagami, J., Tada, N., Miyazaki, S., and Kudo, A. (2000) The gamete fusion process is defective in eggs of CD9-deficient mice. Nat. Genet. 24, 279–282.

    Article  CAS  PubMed  Google Scholar 

  50. Le Naour, F., Rubinstein, E., Jasmin, C., Prenant, M., and Boucheix, C. (2000) Severely reduced female fertility in CD9-deficient mice. Science 287, 319–321.

    Article  CAS  PubMed  Google Scholar 

  51. Miyado, K., Yamada, G., Yamada, S., Hasuwa, H., Nakamura, Y., Ryu, F., Suzuki, K., Kosai, K., Inoue, K., Ogura, A., Okabe, M., and Mekada, E. (2000) Requirement of CD9 on the egg plasma membrane for fertilization. Science 287, 321–324.

    Article  CAS  PubMed  Google Scholar 

  52. Zhu, G. Z., Miller, B. J., Boucheix, C., Rubinstein, E., Liu, C. C., Hynes, R. O., Myles, D. G., and Primakoff, P. (2002) Residues SFQ (173–175) in the large extracellular loop of CD9 are required for gamete fusion. Development 129, 1995–2002.

    CAS  PubMed  Google Scholar 

  53. Chen, M. S., Tung, K. S. K., Coonrod, S. A., Takahashi, Y., Bigler, D., Chang, A., Yamashita, Y., Kincade, P. W., Herr, J. C., and White, J. M. (1999) Role of integrin-associated protein CD9 in binding between sperm ADAM2 and the egg integrin α 6β1: implications for murine fertilization. Proc. Natl. Acad. Sci., U.S.A. 96, 11830–11835.

    Article  CAS  PubMed  Google Scholar 

  54. Ellerman, D. A., Ha, C., Primakoff, P., Myles, D. G., and Dveksler, G. S. (2003) Direct binding of the ligand PSG17 to CD9 requires a CD9 site essential for sperm–egg fusion. Mol. Biol. Cell 14, 5098–5103.

    Article  CAS  PubMed  Google Scholar 

  55. Takahashi, Y., Bigler, D., Ito, Y., and White, J. M. (2001) Sequence specific interaction between the disintegrin domain of mouse ADAM 3 and murine eggs: role of beta1 integrin-associated proteins CD9, CD81, and CD98. Mol. Biol. Cell 12, 809–820.

    CAS  PubMed  Google Scholar 

  56. Inoue, N., Ikawa, M., Isotani, A., and Okabe, M. (2005) The immunoglobin superfamily protein Izumo is required for sperm to fuse with eggs. Nature 434, 234–238.

    Article  CAS  PubMed  Google Scholar 

  57. Ellerman, D. A., Da Ros, V. G., Cohen, D. J., Busso, D., Morgenfeld, M. M., and Cuasnicu, P. S. (2002) Expression and structure–function analysis of de, a sperm cysteine-rich secretory protein that mediates gamete fusion. Biol. Reprod. 67, 1225–1231.

    Article  CAS  PubMed  Google Scholar 

  58. Alfieri, J. A., Martin, A. D., Takeda, J., Kondoh, G., Myles, D. G., and Primakoff, P. (2003) Infertility in female mice with an oocyte-specific knockout of GPI-anchored proteins. J. Cell Sci. 116, 2149–2155.

    Article  CAS  PubMed  Google Scholar 

  59. Coonrod, S. A., Naaby-Hansen, S., Shetty, J., Shibahara, H., Chen, M., White, J. M., and Herr, J. C. (1999) Treatment of mouse oocytes with PI-PLC releases 70-kDa (pI 5) and 35- to 45-kDa (pI 5.5) protein clusters from the egg surface and inhibits sperm–oolemma binding and fusion. Dev. Biol. 207, 334–349.

    Article  CAS  PubMed  Google Scholar 

  60. Kondoh, G., Tojo, H., Nakatani, Y., Komazawa, N., Murata, C., Yamagata, K., Maeda, Y., Kinoshita, T., Okabe, M., Taguchi, R., and Takeda, J. (2005) Angiotensin-converting enzyme is a GPI-anchored protein releasing factor crucial for fertilization. Nat. Med. 11, 160–166.

    Article  CAS  PubMed  Google Scholar 

  61. Fuchs, S., Frenzel, K., Hubert, C., Lyng, R., Muller, L., Michaud, A., Xiao, H. D., Adams, J. W., Capecchi, M. R., Corvol, P., Shur, B. D., and Bernstein, K. E. (2005) Male fertility is dependent on dipeptidase activity of testis ACE. Nat. Med. 11, 1139–1140.

    Article  Google Scholar 

  62. Leisle, L., Parkin, E. T., Turner, A. J., Hooper, N. M. (2005) Angiotensin-convert- ing enzyme as a GPIase: a critical reevaluation. Nat. Med. 11, 1140–1142.

    Article  Google Scholar 

  63. Runft, L. L., Jaffe, L. A., and Mehlmann, L. M. (2002) Egg activation at fertiliza-tion: where it all begins. Dev. Biol. 245, 237–254.

    Article  CAS  PubMed  Google Scholar 

  64. Swann, K. and Jones, K. T. (2002) Membrane events of egg activation, in Fertilization (Hardy, D. M., ed.). Academic Press, San Diego, pp. 319–346.

    Chapter  Google Scholar 

  65. Ducibella, T. (1996) The cortical reaction and development of activation compe- tence in mammalian oocytes. Hum. Reprod. Update 2, 29–42.

    Article  CAS  PubMed  Google Scholar 

  66. Abbott, A. L. and Ducibella, T. (2001) Calcium and the control of mammalian cortical granule exocytosis. Front. Biosci. 6, d792–806.

    Article  CAS  PubMed  Google Scholar 

  67. Tsaadon, A., Eliyahu, E., Shtraizent, N., and Shalgi, R. (2006) When a sperm meets an egg: block to polyspermy. Mol. Cell. Endocrinol. 252, 107–114.

    Article  CAS  PubMed  Google Scholar 

  68. Eliyahu, E., Tsaadon, A., Shtraizent, N., and Shalgi, R. (2005) The involvement of protein kinase C and actin filaments in cortical granule exocytosis in the rat. Reproduction 129, 161–170.

    Article  CAS  PubMed  Google Scholar 

  69. Rossi, E. A., Li, Z., Feng, H., and Rubin, C. S. (1999) Characterization of the targeting, binding, and phosphorylation site domains of an A kinase anchor protein and a myristoylated alanine-rich C kinase substrate-like analog that are encoded by a single gene. J. Biol. Chem. 274, 27201–27210.

    Article  CAS  PubMed  Google Scholar 

  70. Evans, J. P. (1999) Sperm disintegrins, egg integrins, and other cell adhesion molecules of mammalian gamete plasma membrane interactions. Front. Biosci. 4, D114–D131.

    Article  CAS  PubMed  Google Scholar 

  71. Xu, Z., Abbott, A., Kopf, G. S., Schultz, R. M., and Ducibella, T. (1997) Spontaneous activation of ovulated mouse eggs: time-dependent effects on M-phase exit, cortical granule exocytosis, maternal messenger ribonucleic acid recruitment, and inositol 1,4,5-triphosphate sensitivity. Biol. Reprod. 57, 743–750.

    Article  CAS  PubMed  Google Scholar 

Additional References

  • Austin C. R. and Short R. V. (eds.) (1983) Reproduction in Mammals, Book 1, Germ Cells and Fertilization. Cambridge University Press, Cambridge, England.

    Google Scholar 

  • Dunbar B. S. and O'Rand M.G. (eds.) (1991) A Comparative Overview of Mammalian Fertilization. Plenum Press, New York.

    Google Scholar 

  • Gwatkin R. B. L. (1977) Fertilization Mechanisms in Man and Mammals. Plenum Press, New York.

    Google Scholar 

  • Wassarman P. M. (ed.) (1991) Elements of Mammalian Fertilization, vols. 1 and 2. CRC Press, Boca Raton, FL.

    Google Scholar 

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Acknowledgments

We are grateful to Elizabeth Chen for her kind invitation to write this chapter. Whenever possible, we have referred to articles and reviews published between 2000 and 2006. Our laboratory is supported in part by the National Institutes of Health (HD35105).

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Authors and Affiliations

  1. Department of Molecular, Cell and Developmental Biology, Mount Sinai School of Medicine, New York, NY

    Paul M. Wassarman & Eveline S. Litscher

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  1. Paul M. Wassarman
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Editors and Affiliations

  1. Department of Molecular Biology and Genetics, Johns Hopkins University School of Medicine, Baltimore, MD

    Elizabeth H. Chen

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Wassarman, P.M., Litscher, E.S. (2008). Mammalian Fertilization Is Dependent on Multiple Membrane Fusion Events*. In: Chen, E.H. (eds) Cell Fusion. Methods in Molecular Biology™, vol 475. Humana Press. https://doi.org/10.1007/978-1-59745-250-2_6

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  • DOI: https://doi.org/10.1007/978-1-59745-250-2_6

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-911-6

  • Online ISBN: 978-1-59745-250-2

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