Hostname: page-component-586b7cd67f-t8hqh Total loading time: 0 Render date: 2024-11-26T14:09:44.007Z Has data issue: false hasContentIssue false

Unprotected freezing of human spermatozoa exerts a detrimental effect on their oocyte activating capacity and chromosome integrity

Published online by Cambridge University Press:  26 September 2008

Andrei V. Rybouchkin*
Affiliation:
Infertility Centre, Department of Gynaecology and Obstetrics, University Hospital of Ghent, Ghent, Belgium.
Paul De Sutter
Affiliation:
Infertility Centre, Department of Gynaecology and Obstetrics, University Hospital of Ghent, Ghent, Belgium.
Marc Dhont
Affiliation:
Infertility Centre, Department of Gynaecology and Obstetrics, University Hospital of Ghent, Ghent, Belgium.
*
Andrei V. Rybouchkin, Infertility Centre, Department of Gynaecology and Obstetrics, University Hospital, De Pinetelaan 185, B-9000 Ghent, Belgium. Telephone +32 9 2402194. Fax: +32 9 240 4972. e-mail: [email protected].

Summary

The influence of unprotected freezing of mammalian spermatozoa on their oocyte activating capacity and chromosome integrity is unknown. However, this type of sperm treatment has been used in assisted reproduction by intracytoplasmic sperm injection in cattle and humans. The mouse oocyte injection test was used to analyse the influence of unprotected freezing of human spermatozoa on their reproductive characteristics. Mouse oocytes were microinjected with intact human spermatozoa or spermatozoa treated with two cycles of unprotected freeze-thawing. Oocytes surviving the injection were either cultured without further treatment or exposed to ethanol solution to induce parthenogenetic activation. Both injected and activated oocytes were used for sperm chromosome analysis. The results revealed a significant reduction in oocyte activating capacity and a tenfold increase in the incidence of structural chromosomal abnormalities in human spermatozoa treated by unprotected freezing. We conclude that unprotected freezing of human spermatozoa has a detrimental effect on their reproductive characteristics. Our data also provide a new perspective on the stability of mammalian spermatozoa to physical factors and demonstrate the importance of detailed analysis of the stability of sperm structures for successful development of new approaches in assisted reproduction.

Type
Article
Copyright
Copyright © Cambridge University Press 1996

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

Boerian, M.L. & Saris, L.A. (1991). The effect of spermatozoal irradiation woth X-rays on chromosome abnomalities and on development of mouse zygotes after delayed fertilization. Mutation Res. 256, 4957.CrossRefGoogle Scholar
Dozortsev, D., Rybouchkin, A., De Sutter, P., Qian, C. & Dhont, M. (1995). Human oocyte activation following intracytoplasmic injection: the role of the sperm cell. Hum. Reprod. 10, 403–7.CrossRefGoogle ScholarPubMed
Dyban, A. (1983). An improved method for chromosome preparation from preimplantation mammalian embryos, oocytes or isolated blastomeres. Stain Technol. 58, 6972.CrossRefGoogle ScholarPubMed
Fulton, B.P. & Whittingham, D.C. (1978). Activation of mammalian oocytes by intracellular injection of calcium. Nature. 273, 149–51.CrossRefGoogle ScholarPubMed
Goto, K. (1993). Bovine microfertilization and embryo transfer. Mol. Reprod. Dev. 36, 288–90.CrossRefGoogle ScholarPubMed
Goto, K., Kinoshita, A. & Takuma, Y. (1990). Fertilization of bovine oocytes by the injection of immobilized, killed spermatozoa. Vet. Rec. 127, 517–20.Google ScholarPubMed
Homa, S.T. & Swann, K. (1994). A cytosolic sperm factor triggers calcium oscillations and membrane hyperpolarizations in human oocytes. Hum. Reprod. 9, 2356–61.CrossRefGoogle ScholarPubMed
Hoshi, K., Yanagida, K. & Sato, A. (1992). Pretreatment of hamster oocytes with Ca2+ ionophore to facilitate fertilization by ooplasmic micro-injection. Hum. Reprod. 7, 871–5.CrossRefGoogle ScholarPubMed
Hoshi, K., Yanagida, K., Katayose, H. & Yazawa, H. (1994 a). Pronuclear formation and cleavage of mammalian eggs after microsurgical injection of freeze-dried sperm nuclei. Zygote. 2, 237–42.CrossRefGoogle ScholarPubMed
Hoshi, K., Yanagida, K., Yazawa, H., Katayose, H. & Sato, A. (1994 b). Pregnancy and delivery after intracytoplasmic injection of an immobilized, killed spermatozoon into an oocyte. J. Assist. Reprod. Genet. 11, 325–6.CrossRefGoogle ScholarPubMed
Hoshi, K., Yanagida, K., Yazawa, H., Katayose, H. & Sato, A. (1995). lntracytoplasmic sperm injection using immobilized, or motile human spermatozoon. Fertil. Steril. 63, 1241–5.CrossRefGoogle ScholarPubMed
ISCN (1985). An international system for human cytogenetic nomenclature. Birth Defects 21, 1117.Google Scholar
Katayose, H., Matsuda, J. & Yanagimachi, R. (1992). The ability of dehydrated hamster and human sperm nuclei to develop into pronuclei. Biol. Reprod. 47, 277–84.CrossRefGoogle ScholarPubMed
Kimura, Y. & Yanagimachi, R. (1995). Intracytoplasmic sperm injection in the mouse. Blot. Reprod. 52, 709–20.CrossRefGoogle ScholarPubMed
Lanzendorf, S.E., Maloney, M.K., Veeck, L.L., Slusser, J., Hodgen, G.D. & Rosenwaks, Z. (1988). A preclinical evaluation of pronuclear formation by microinjection of human spermatozoa into human oocytes. Fertil. Steril. 49, 835–42.Google ScholarPubMed
Martin, R.H., Ko, E. & Rademaker, A. (1988). Human sperm chromosome complements after microinjection of hamster eggs. J. Reprod. Fertil. 84, 179–86.CrossRefGoogle ScholarPubMed
McKiernan, S.H. & Bavister, B.D. (1994). Timing of development is a critical parameter for predicting successful embryogenesis. Hum. Reprod. 9, 2123–9.CrossRefGoogle ScholarPubMed
Munne, S. & Estop, A. (1991). The effect of in vitro ageing on mouse sperm chromosomes. Hum. Reprod. 6, 703–8.CrossRefGoogle ScholarPubMed
Munne, S. & Estop, A. (1993). Chromosome analysis of human spermatozoa stored in vitro. Hum. Reprod. 8, 581–6.CrossRefGoogle ScholarPubMed
Parrington, J., Swann, K., Shevchenko, V.I., Sesay, A.K. & Lai, F.A. (1996). Calcium oscillations in mammalian eggs triggered by a soluble sperm protein. Nature 379, 364–8.CrossRefGoogle ScholarPubMed
Poe-Zeigler, P., Boyd, C.A., Nehchiri, F., Neely, B., Hammacher, P. & Lanzendorf, S.E. (1995). Effect of sperm viability on fertilization and cleavage following intracytoplasmic sperm injection. In Abstracts of the Ffty-first Annual Meeting of the American Society for Reproductive Medicine, 712 October, Seattle, Washington, P-032, S11O. Birmingham, AL: American Society for Reproductive Medicine.Google Scholar
Rybouchkin, A., Dozortsev, D., De Sutter, P., Qian, C. & Dhont, M. (1995). Intracytoplasmic injection of human sperm into mouse oocytes: a useful model to investigate oocyte activating capacity and the karyotype of human sperm. Hum. Reprod. 10, 1130–5.CrossRefGoogle Scholar
Swann, K. (1990). A cytosolic sperm factor stimulates repetitive calcium increases and mimics fertilization in hamster eggs. Development 110, 1295–302.CrossRefGoogle ScholarPubMed
Uehara, T. & Yanagimachi, R. (1976). Microsurgical injection of spermatozoa into hamster eggs with subsequent transformation of sperm nuclei into male pronuclei. Biol. Reprod. 15, 457–70.CrossRefGoogle ScholarPubMed
Uehara, T. & Yanagimachi, R.. (1977). Activation of hamster eggs by pricking. J. Exp. Zool. 199, 269–74.CrossRefGoogle ScholarPubMed
Yadav, B.R., King, W.A. & Betteridge, K.J. (1993). Relationships between the completion of first cleavage and the chromosomal complement, sex, and developmental rates of bovine embryos generated in vitro. Mol. Reprod. Dev. 36, 434–9.CrossRefGoogle ScholarPubMed
Yanagida, K., Yanagimachi, R., Perreault, S.D & Kleinfeld, R.G. (1991). Thermostability of sperm nuclei assessed by microinjection into hamster oocytes. Biol. Reprod. 44, 440–7.CrossRefGoogle ScholarPubMed
Yanagimachi, R.. (1995). Is an animal model needed for intracytoplasmic sperm injection (ICSI) and other assisted reproduction technologies? Hum. Reprod. 10, 2525–6.CrossRefGoogle ScholarPubMed