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Pronuclear formation and intracellular glutathione content of in vitro-matured porcine oocytes following in vitro fertilisation and/or electrical activation

Published online by Cambridge University Press:  26 September 2008

Hiroaki Funahashi ,
Todd T. Stumpf ,
Thomas C. Cantley ,
Nam-Hyung Kim  and
Billy N. Day
Hiroaki Funahashi
Affiliation:
Department of Animal Sciences, University of Missouri-Columbia, Columbia, Missouri USA
Todd T. Stumpf
Affiliation:
Department of Animal Sciences, University of Missouri-Columbia, Columbia, Missouri USA
Thomas C. Cantley
Affiliation:
Department of Animal Sciences, University of Missouri-Columbia, Columbia, Missouri USA
Nam-Hyung Kim
Affiliation:
Department of Animal Sciences, University of Missouri-Columbia, Columbia, Missouri USA
Billy N. Day*
Affiliation:
Department of Animal Sciences, University of Missouri-Columbia, Columbia, Missouri USA
*
Billy N. Day, University of Missouri Columbia, Department of Animal Sciences, 159 Animal Sciences Center, Columbia, MO 65211, USA. Telephone: (314) 882-7555. Fax: (314) 882-6827.
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Summary

Pronuclear formation and intracellular content of glutathione, containing reduced and oxidised forms, in porcine oocytes matured in vitro were determined following insemination and/or electrical stimulation. After insemination, sperm penetration had occurred as early as 3 h and female pronuclei had formed by 6 h with complete development by 12 h. Male pronuclear formation occurred, primarily, between 9 and 12 h after insemination. Glutathione content of the oocytes decreased following sperm penetration and remained at a depressed level until 12 h. After electrical stimulation, oocyte activation had occurred and female pronuclei had formed by 3 and 6 h, respectively. Oocyte glutathione content did not change as a result of oocyte activation. When oocytes were exposed to an electrical pulse and then spermatozoa, female pronuclear formation was observed by 3 h after stimulation/insemination. Sperm penetration was observed between 3 and 9 h. However, the incidence of male pronuclear formation observed at 12 h was extremely low, although sperm decondensation had occurred in some oocytes. Oocyte glutathione content had not decreased by 6 h following electrical activation. These results demonstrate that the changes in glutathione content in porcine oocytes following fertilisation in vitro differ from those due to electrical activation. Further, the decreased intracellular glutathione content in oocytes activated by sperm penetration appears to be due to the presence of a sperm factor.

Keywords

FertilisationGlutathioneOocytePigPronuclear formation

Information

Type
Article
Information
Zygote , Volume 3 , Issue 3 , August 1995 , pp. 273 - 281
Copyright
Copyright © Cambridge University Press 1995

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References

Akerboom, T. & Sies, H. (1990). Glutathione transport and its significance in oxidative stress. In Glutathione: Metabolism and Physiological Functions, ed. Vina, J.. 45–55. Boca Raton, FL: CRC Press.Google Scholar
Anderson, M.E. (1985). Determination of glutathione and glutathione disulfide in biological samples. Methods Enzymol 113, 548–55.Google ScholarPubMed
Beckmann, L.S. & Day, B.N. (1993). Effect of media NaCl concentration and osmolarity on culture of the early stage porcine embryo and viability of embryos cultured in a selected superior medium. Theriogenology 39, 611–22.CrossRefGoogle Scholar
Chance, B., Sies, H. & Boveris, A. (1979). Hydroperoxide metabolism in mammalian organs. Physiol. Rev. 59, 527–605.CrossRefGoogle ScholarPubMed
Cran, D.G. & Cheng, W.T.K. (1986). The cortical reaction in pig oocytes during in vivo and in vitro fertilization. Gamete Res. 13, 241–51.CrossRefGoogle Scholar
Ding, J., Clarke, N., Nagai, T. & Moor, R.M. (1992). Protein and nuclear changes in pig eggs at fertilisation. Mol. Reprod. Dev. 31, 287–96.CrossRefGoogle Scholar
Funahashi, H. & Day, B.N. (1993a). Effects of different serum supplements in maturation medium on meiotic and cytoplasmic maturation of pig oocytes. Theriogenology 39, 965–73.CrossRefGoogle ScholarPubMed
Funahashi, H. & Day, B.N. (1993b). Effects of follicular fluid at fertilisation in vitro on sperm penetration in pig oocytes. J. Reprod. Fertil. 99, 97–103.CrossRefGoogle ScholarPubMed
Funahashi, H. & Day, B.N. (1993c). Effects of the duration of exposure to supplemental hormones on cytoplasmic maturation of pig oocytes in vitro. J. Reprod. Fertil. 98, 179–85.CrossRefGoogle Scholar
Funahashi, H., Stumpf, T.T., Terlouw, S.L. & Day, B.N. (1993). Effects of electrical stimulation before or after in vitro fertilization on sperm penetration and pronuclear formation in pig oocytes. Mol. Reprod. Dev. 36, 361–7.CrossRefGoogle ScholarPubMed
Funahashi, H., Cantley, T.C. & Day, B.N. (1994a). Different hormonal requirement of porcine oocyte-complexes during maturation in vitro. J. Reprod. Fert. 101, 159–65.CrossRefGoogle Scholar
Funahashi, H., Cantley, T.C., Stumpf, T.T., Terlouw, S.L. & Day, B.N. (1994b). In vitro development of in vitro matured porcine oocytes following chemical activation or in vitro fertilization. Biol. Reprod. 50, 1072–77.CrossRefGoogle ScholarPubMed
Funahashi, H., Cantley, T.C., Stumpf, T.T., Terlouw, S.L. & Day, B.N. (1994c). Use of low salt culture medium for in vitro maturation of porcine oocytes is associated with elevated oocyte glutathione levels and enhanced male pronuclear formation after in vitro fertilization. Biol. Reprod. 51, 633–9.CrossRefGoogle ScholarPubMed
Funahashi, H., Stumpf, T.T., Terlouw, S.L., Cantley, T.C., Rieke, A. & Day, B.N. (1994d). Developmental ability of porcine oocytes matured and fertilized in vitro. Theriogenology 41, 1425–33.CrossRefGoogle ScholarPubMed
Gardiner, C.S. & Reed, D.J. (1994). Status of glutathione during oxidant-induced oxidative stress in the preimplantation mouse embryo. Biol. Reprod. 51, 1307–14.CrossRefGoogle ScholarPubMed
Hunter, R.H.F. (1972). Fertilization in the pig: sequence of nuclear and cytoplasmic events. J. Reprod. Fertil. 29, 395–406.CrossRefGoogle ScholarPubMed
Hunter, R.H.F. (1974). Chronological and cytological details of fertilization and early embryonic development in the domestic pig, Sus scrofa. Anat. Rec. 178, 169–86.CrossRefGoogle ScholarPubMed
Hunter, R.H.F. (1976). Sperm—egg interactions in the pig: monospermy, extensive polyspermy, and the formation of chromatin aggregates. J. Anat. 122, 43–59.Google ScholarPubMed
Kosower, N.S. (1978). The glutathione status of cells. Inter. Rev. Cytol. 54, 109–60.CrossRefGoogle ScholarPubMed
Meister, A. & Anderson, M.E. (1983). Glutathione. Annu. Rev. Biochem. 52, 11–60.CrossRefGoogle ScholarPubMed
Naito, K. & Toyoda, Y. (1992). Effects of microinjection of glutathione on male pronucleus formation in porcine oocytes matured in vitro. J. Reprod. Dev. 38, 173–8.CrossRefGoogle Scholar
Perreault, S.D. (1990). Regulation of sperm nuclear reactivation during fertilization. In Fertilization in Mammals, ed. Bavister, B.D., Cummins, J. & Roldan, E.R.S. pp. 285–96. Norwell, MA: Serono Symposia, USA.Google Scholar
Perreault, S.D., Barbee, R.R. & Slott, V.L. (1988). Importance of glutathione in the acquisition and maintenance of sperm nuclear decondensing activity in maturing hamster oocytes. Dev. Biol. 125, 181–6.CrossRefGoogle ScholarPubMed
Prather, R.S., Eichen, P.A., Nicks, D.K. & Peters, M.S. (1991). Artificial activation of porcine oocytes matured in vitro. Mol. Reprod. Dev. 28, 405–9.CrossRefGoogle ScholarPubMed
Sun, F.Z., Hoyland, J., Huang, X., Mason, W. & Moor, R.M. (1992). A comparison of intercellular changes in porcine eggs after fertilization and electroactivation. Development 115, 947–56.CrossRefGoogle Scholar
Whitten, W.K. & Biggers, J.D. (1968). Complete development in vitro of the pre-implantation stages of the mouse in a simple chemically defined medium. J. Reprod. Fertil. 17, 399–401.CrossRefGoogle Scholar
Yanagimachi, R. (1981). Mechanisms of fertilization in mammals. In Fertilization and Embryonic Development In Vitro, ed. Mastroanni, L.J. & Biggers, J.D. pp. 81–182. New York: Plenum Press.CrossRefGoogle Scholar
Yanagimachi, R. (1994). Mammalian fertilization. In The Physiology of Reproduction, ed. Knobil, E., Neill, J.D., Greenwald, G.S., Markert, C.L. & Pfaff, D.W. pp. 189–317. New York: Raven Press.Google Scholar
Yoshida, M. (1993). Role of glutathione in the maturation and fertilization of pig oocytes in vitro. Mol. Reprod. Dev. 35, 76–81.CrossRefGoogle ScholarPubMed
Yoshida, M., Cran, D.G. & Pursel, V.G. (1993a). Confocal and fluorescence microscopic study using lectins of the distribution of cortical granules during the maturation and fertilization of pig oocytes. Mol. Reprod. Dev. 36, 462–8.CrossRefGoogle ScholarPubMed
Yoshida, M., Ishigaki, K., Nagai, T., Chikyu, M. & Pursel, V.G. (1993b). Glutathione concentration during maturation and after fertilization in pig oocytes: relevance to the ability of oocytes to form male pronucleus. Biol. Reprod. 49, 89–94.CrossRefGoogle Scholar
Zirkin, B.R., Soucek, D.A., Chang, T.S.K. & Perreault, S.D. (1985). In vitro and in vivo studies of mammalian sperm nuclear decondensation. Gamete Res. 11, 349–65.CrossRefGoogle Scholar
Zirkin, B.R., Perreault, S.D. & Naish, S.J. (1989). Formation and function of the male pronucleus during mammalian fertilization. In The Molecular Biology of Fertilization, ed. Schatten, H. & Schatten, G. pp. 91–114. San Diego, CA: Academic Press.CrossRefGoogle Scholar