Hostname: page-component-78c5997874-lj6df Total loading time: 0 Render date: 2024-11-02T19:03:56.426Z Has data issue: false hasContentIssue false
  • English
  • Français

Mouse sperm fertilising capacity following subzonal microinjection is dependent on sperm washing and response to solubilised zonae pellucidae

Published online by Cambridge University Press:  26 September 2008

Orly Lacham Kaplan  and
Alan Trounson
Orly Lacham Kaplan
Affiliation:
Institute for Reproduction and Development, Monash University, Melbourne, Australia.
Alan Trounson*
Affiliation:
Institute for Reproduction and Development, Monash University, Melbourne, Australia.
*
Dr Alan Trounson, Centre for Early Human Development, Monash University, Monash Medical Centre, 246 Clayton Road, Clayton, Victoria 3168, Australia. Telephone: 61-3-550-5300. Fax: 61-3-550 5554.
Get access Rights & Permissions [Opens in a new window]

Summary

Spermatozoa obtained from hybrid male mice were treated with solubilised zonae pellucidae after a period of capacitation in vitro to induce the acrosome reaction. Single spermatozoa were selected and microinjected into the perivitelline space of mature oocytes. A high proportion of the spermatozoa acrosome-reacted affter treatment with solubilised zonae (63%). However, the fertilisation rate (37%) after subzonal microinjection of oocytes by the treated spermatozoa was not different to the fertilisation rate (40%) of oocytes microinjected with untreated spermatozoa which had a lower rate of acrosome reaction (39%). When spermatozoa were washed by high-speed centrifugation before treatment with solubilised zonae pellucidae and subzonal microinjection, the fertilisation rate (68%) was significantly higher (p < 0.001) than that (29%) for occytes microinjected with untreated spermatozoa and was found to be correlated with a high acrosome reaction rate (74%) (r = 0.8). The washing of spermatozoa by centrifugation itself did not increase the acrosome reaction rate or fertilisation rate of oocytes after microinjection. The results of this study suggests that some modifications other than the acrosome reaction are needed to enable capacitated mouse spermatozoa to fuse with the oocyte plasma membrane. These modifications were achieved by washing spermatozoa by high-speed centrifugation and the replacement of the supernatant with fresh culture medium used for capacitation. Induction of the acrosome reaction by solubilised zonae pellucidae following this treatment leads to a high fertilisation rate of oocytes by subzonal sperm microinjection.

Keywords

Acrosome reactionFertilisationMicroinjectionSpermatozoaZona pellucida
Type
Article
Information
Zygote , Volume 3 , Issue 1 , February 1995 , pp. 9 - 16
Copyright
Copyright © Cambridge University Press 1995

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

Aarons, D., Speake, J.L., & Poirier, R.G.. (1984). Evidence for a proteinase inhibitor binding component associated with murine spermatozoa. Biol. Reprod. 31, 811–17.CrossRefGoogle ScholarPubMed
Aarons, D., Battle, T., Boettger-Tong, H., & Poirier, G.R. (1992). Role of monoclonal antibody J-23 in inducing acrosome reaction in capacitated mouse spermatozoa. J. Reprod. Fert. 96, 49–59.CrossRefGoogle ScholarPubMed
Battle, T.A., Boettger, H.L., & Poirier, G.R.. (1990). An epididymal component capable of masking a sperm head epitope. Biol. Reprod. 42, Suppl. 1, 111 (abstr.).Google Scholar
Bleil, J.D., & Wassarman, P.M.. (1980). Structure and function of the zona pellucida: identification and characterization of the proteins of the mouse oocyte's zona pellucida. Dev. Biol. 76, 185–202.CrossRefGoogle ScholarPubMed
Bleil, J.D., & Wassarman, P.M.. (1983). Sperm–egg interaction in the mouse: sequence of events and induction of the acrosome reaction by a zona pellucida glycoprotein. Dev. Biol. 95, 317–24.CrossRefGoogle ScholarPubMed
Bleil, J.D., Greve, J.M., & Wassarman, P.M.. (1988). Induction of a a secondary sperm receptor in the mouse egg zona pellucida: role in maintenance of binding of acrosome reacted sperm. Dev. Biol. 128, 376–85.CrossRefGoogle Scholar
Bryan, J.H.D.. (1970). An eosin fat green naphthol yellow mixture for differential staining of cytologic components in mammalian spematozoa. Stain Technol. 45, 231–6.CrossRefGoogle Scholar
Florman, H.M., Bechtol, K.B., & Wassarman, P.M.. (1984). Enzymatic dissection of the functions of the mouse egg's receptor for sperm. Dev. Biol. 106, 243–55.CrossRefGoogle ScholarPubMed
Fraser, L.R.. (1983). Mouse sperm capacitation assessed by kinetics and morphology of fertilization in vitro. J. Reprod. Fert. 69, 539–53.CrossRefGoogle ScholarPubMed
Fraser, L.R.. (1984). Mouse sperm capacitation in vitro involves loss of a surface-associated inhibitory component. J. Reprod. Fert. 72, 373–84.CrossRefGoogle ScholarPubMed
Fraser, L.R., Harrison, R.A.P., & Herod, J.E.. (1990). Characterization of a decapacitation factor associated with epididymal mouse spermatozoa. J. Reprod. Fert. 89, 135–48.CrossRefGoogle ScholarPubMed
Lacham, O., & Trounson, A.. (1991). Fertilizing capacity of epididymal and testicular spermatozoa microinjected under the zona pellucida of mouse oocytes. Mol. Reprod. Dev. 29, 85–93.CrossRefGoogle Scholar
Lacham, O., Trounson, A., Holdern, C., Mann, J., & Sathananthan, H.. (1989). Fertilization and development of mouse eggs injected under the zona pellucida with single spermatozoa treated to induce the acrosome reaction. Gamete Res. 23, 233–43.CrossRefGoogle ScholarPubMed
Lopez, L.C., & Shur, B.D.. (1987). Redistribution of mouse sperm surface galactosyltransferase after the acrosome reaction. J. Cell Biol.. 105, 1663–70.CrossRefGoogle ScholarPubMed
Lopez, L.C., Bayna, E.M., Litoff, D., Shaper, N.L., Shaper, J.H., & Shur, B.D.. (1985). Receptor function of mouse sperm surface galactosyltransferase during ferilization. J. Cell Biol. 101, 1501–10.CrossRefGoogle Scholar
Mann, J.R.. (1988). Full term development of mouse eggs fertilized by a spermatozoon microinjected under the zona pellucida. Biol. Reprod. 38, 1077–83.CrossRefGoogle ScholarPubMed
Miller, D.J., Macek, M.B., & Shur, B.D.. (1992). Complementarity between sperm surface β-1, 4-galactosyl-transferase and egg coat ZP3 mediates sperm–egg binding. Nature 357, 589–93.CrossRefGoogle Scholar
Myles, D.G., & Primakoff, P.. (1984). Localized surface antigens of guinea pig sperm migrate to new regions prior to fertilization. J. Cell Biol. 99, 1634–41.CrossRefGoogle ScholarPubMed
Quinn, P., Barros, C., & Whittingham, D.G.. (1982). Preservation of hamster oocytes to assay the fertilizing capacity of human spermatozoa. J. Reprod. Fert. 66. 161–8.CrossRefGoogle ScholarPubMed
Saling, P.M., Irons, G., & Waibel, R.. (1985). Mouse sperm antigens that participate in fertilisation. I. Inhibition of sperm fusion with the egg plasma membrane using monoclonal antibodies. Biol. Reprod. 33, 515–26.CrossRefGoogle ScholarPubMed
Saxena, N.K., Russel, L.D., Saxena, N., & Peterson, R.N.. (1986). Immunofluorescence antigen localization on boar sperm plasma membranes: monoclonal antibodies reveal apparent new domains and apparent redistribution of surface antigens during sperm maturation and ejaculation. Anat. Rec. 214, 238–52.CrossRefGoogle ScholarPubMed
Shapiro, B.M., Schackmann, R.W., & Gabel, C.A.. (1981). Molecular approaches to the study of fertilization. Annu. Rev. Biochem. 50, 815–43.CrossRefGoogle Scholar
Shur, B.D., & Hall, N.G.. (1982). Sperm surface galactosyltransferase activities during in vitro capacitation. J. Cell Biol. 95, 567–73.CrossRefGoogle ScholarPubMed
Soldani, P., & Rosati, F.. (1987). Sperm-egg interaction in the mouse using live and glutaraldehyde fixed eggs. Gamete Res. 18, 225–35.CrossRefGoogle ScholarPubMed
Storey, B.T., Lee, M.A., Muller, L., Ward, C.R., & Wirtchafter, D.G.. (1984). Binding of mouse spermatozoa to the zonae pellucidae of mouse eggs in cumulus: evidence that the acrosome remain substantially intact. Biol. Reprod. 31, 1119–28.CrossRefGoogle Scholar
Villarroya, S., & Scholler, R.. (1987). Lateral diffusion of a human sperm head antigen during incubation in a capacitated medium and induction of the acrosome reaction in vitro. J. Reprod. Fert. 80. 545–62.CrossRefGoogle Scholar
Wasserman, P.M.. (1990). Profile of a mammalian sperm receptor. Development 108, 1–17.CrossRefGoogle Scholar
Wasserman, P.M., Bleil, J.D., Florman, J.M., Greve, J.M., Roller, R.J., Salzmann, G.S., & Samuels, F.G.. (1989). The mouse egg's receptor for sperm: a multifunctional zona pellucida glycoprotein. In The Mammalian Egg Coat: Structure and Function, ed. Dietl, J, pp 18–37. Berlin: Springer.CrossRefGoogle Scholar
Whittingham, G.D.. (1971). Culture of mouse ova. J. Reprod. Fert. Suppl. 14, 7–21.Google ScholarPubMed
Yanagimachi, R.. (1988). Mammalian fertilization. In The Physiology of Reproduction, vol. 1, ed. Knobil, E., & Neill, J.D., 135–85. New York: Raven Press.Google Scholar