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Transformation of sperm nuclei into metaphase chromosomes in maturing pig oocytes penetrated in vitro

Published online by Cambridge University Press:  26 September 2008

Wei-Hua Wang  and
Koji Niwa
Wei-Hua Wang
Affiliation:
Division of Animal Science and Technology, Faculty of Agriculture, Okayama University, Okayama 700, Japan
Koji Niwa*
Affiliation:
Division of Animal Science and Technology, Faculty of Agriculture, Okayama University, Okayama 700, Japan
*
Koji Niwa, Division of Animal Science and Technology, Faculty of Agriculture, Okayama University, 1-1-1 Tsushima-Naka, Okayama 700, Japan. Telephone: +81(0)86-251-8328. Fax: +81(0)86-254-0714. e-mail: [email protected].
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Summary

Cumulus-free pig oocytes at the germinal vesicle (GV) stage were incubated in modified Brackett & Oliphant's medium with 5% fetal calf serum and 5mM caffeine with or without cryopreserved, ejaculated spermatozoa. When oocytes were transferred into modified tissue culture medium (TCM- 199B at pH 7.4) supplemented with 1OIU/ml eCG, 1OIU/ml hCG and 1 μg/ml oestradiol-17p after 8h of incubation with spermatozoa and cultured for 0–48 h, 86–99% of oocytes were penetrated. Most (95–100%) oocytes penetrated 0–16 h after transfer had decondensed sperm chromatin. However, 24 h after transfer 47% and 33% of penetrated oocytes contained recondensed sperm chromatin and sperm metaphase chromosomes, respectively. The proportion of penetrated oocytes containing sperm metaphase chromosomes increased after 36–48 h of transfer (51–65%). The transformation of sperm nuclei to metaphase chromosomes was obtained in 75% and 79% of anaphase I (AI) to telophase I (TI) and metaphase II (Mil) oocytes, respectively, but only in 38% of metaphase I (MI) oocytes. Moreover, such transformation was observed only in 1 of 30 oocytes at the stages of GV breakdown to prometaphase I and none of 69 oocytes at the GV stage. The transformation of sperm nuclei into metaphase chromosomes was completely inhibited in oocytes penetrated by eight or more spermatozoa. Well-developed male and female pronuclei were observed in only 3 (4%) of 77 oocytes penetrated 48 h after transfer. The proportion of oocytes reaching Mil was greatly inhibited by sperm penetration; only 18% of penetrated oocytes, but 87% of non-inseminated oocytes, reached Mil by 48 h after transfer. None of the oocytes penetrated by seven or more spermatozoa reached MIL Most (75%) oocytes were inhibited from the transition from MI to Mil even though they were cultured for 48 h. The present results indicate that: (1) the cytoplasm of maturing oocytes possesses an activity for transforming sperm nuclei into metaphase chromosomes, (2) immature pig oocytes penetrated by spermatozoa can undergo meiotic maturation to MI, and (3) the transition of such oocytes from MI to Mil is inhibited, suggesting that an activity of mitogen-activated protein kinase may be retarded.

Keywords

In vitroOocytesPigSperm metaphase chromosomesSperm penetration
Type
Article
Information
Zygote , Volume 5 , Issue 2 , May 1997 , pp. 183 - 191
Copyright
Copyright © Cambridge University Press 1997

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References

Abeydeera, L.R. & Niwa, K. (1992). Ability of in vitro maturing bovine oocytes to transform sperm nuclei to metaphase chromosomes. J. Reprod. Fertil. 96, 565–72.CrossRefGoogle ScholarPubMed
Abeydeera, L.R., Niwa, K. & Okuda, K. (1993). Maturation-promoting factor (MPF) is responsible for the transformation of sperm nuclei to metaphase chromosomes in maturing bovine oocytes in vitro. J. Reprod. Fertil. 98, 409–14.CrossRefGoogle ScholarPubMed
Abeydeera, L.R., Okuda, K. & Niwa, K. (1994). Activation of bovine oocytes penetrated after germinal vesicle breakdown. Zygote 2, 273–9.CrossRefGoogle ScholarPubMed
Balakier, H. (1978). Induction of maturation in small oocytes from sexually immature mice by fusion with meiotic and mitotic cells. Exp. Cell Res. 112, 137–41.CrossRefGoogle ScholarPubMed
Barros, C. & Munoz, G. (1974). Sperm penetration through the zona pellucida of immature hamster oocytes. Acta Physiol. Latinoam. 24, 612–15.Google ScholarPubMed
Brackett, B.G. & Oliphant, G. (1975). Capacitation of rabbit spermatozoa in vitro. Biol Reprod. 12, 260–74.CrossRefGoogle ScholarPubMed
Calvin, H.I., Grosshans, K. & Blake, E.J. (1986). Estimation and manipulation of glutathione levels in prepubertal mouse ovaries and ova: relevance to sperm nucleus transformations in the fertilized egg. Gamete Res. 14, 265–75.CrossRefGoogle Scholar
Clarke, H.J. & Masui, Y. (1985). Inhibition by dibutyrylcyclic AMP of the transition to metaphase of mouse oocyte nuclei and its reversal by cell fusion to metaphase oocytes. Deo. Biol. 108, 32–7.CrossRefGoogle Scholar
Clarke, H.J. & Masui, Y. (1986). Transformation of sperm nuclei to metaphase chromosomes in the cytoplasm of maturing oocytes of the mouse. J. Cell Biol. 102, 1039–46.CrossRefGoogle ScholarPubMed
Das, N.H. & Barker, C. (1976). Mitotic chromosome condensation in the sperm nucleus during postfertilization maturation division in Urechis eggs. J. Cell Biol. 68, 155–9.CrossRefGoogle ScholarPubMed
Elinson, R.P. (1977). Fertilization of immature frog eggs: cleavage and development following subsequent activation. J. Embryol. Exp. Morphol. 37, 187–201.Google ScholarPubMed
Fulka, J., JrFlechon, J.-E., Motilik, J. & Fulka, J. (1988). Does autocatalytic amplification of maturation promoting factor (MPF) exist in mammalian oocytes? Gamete Res. 21, 185–92.CrossRefGoogle ScholarPubMed
Gotoh, Y., Moriyama, K., Matsuda, S., Okumura, E., Kishi-moto, T., Kawasaki, H., Suzuku, K., Yahara, I., Sakai, H. & Nishida, E. (1991). Xenopus M phase MAP kinase: isolation of its cDNA and activation by MPF. EMBO J. 10, 2661–8.CrossRefGoogle ScholarPubMed
Hashimoto, N. & Kishimoto, T. (1988). Regulation of meiotic metaphase by a cytoplasmic maturation promoting factor during mouse oocyte maturation. Dev. Biol. 126, 242–52.CrossRefGoogle ScholarPubMed
Iwamatsu, T. & Chang, M.C. (1972). Sperm penetration in vitro of mouse oocytes at various times after maturation. J. Reprod. Fertil. 31, 237–47.CrossRefGoogle Scholar
Kishimoto, T. & Kanatani, H. (1976). Cytoplasmic factor responsible for germinal vesicle breakdown and meiotic maturation in starfish oocytes. Nature 260, 321–2.CrossRefGoogle Scholar
Kubiak, J.Z., Weber, M., Geraud, G. & Maro, B. (1992). Cell cycle modification during the transition between meiotic M-phases in mouse oocytes. J. Cell Sci. 102, 456–67.CrossRefGoogle ScholarPubMed
Lohka, M.J. (1989). Mitotic control by metaphase-promoting factor and cdc proteins. J. Cell Sci. 92, 131–5.CrossRefGoogle ScholarPubMed
Longo, F.J., Cook, S. & Mathews, L. (1991). Pronuclear formation in starfish eggs inseminated at different stages of meiotic maturation: correlation of sperm nuclear transformations and activity of the maternal chromatin. Dev. Biol. 147, 62–72.CrossRefGoogle ScholarPubMed
Mahi, C.A. & Yanagimachi, R. (1876). Maturation and sperm penetration of canine ovarian oocytes in vitro. J. Exp. Zool. 196, 189–96.CrossRefGoogle Scholar
Masui, Y. & Clarke, H.J. (1979). Oocyte maturation. Int. Rev. Cytol. 57, 185–282.CrossRefGoogle ScholarPubMed
Masui, Y. & Markert, C.L. (1971). Cytoplasmic control of nuclear behaviour during meiotic maturation of frog oocytes. J. Exp. Zool. XT7, 129–46.CrossRefGoogle ScholarPubMed
Mattioli, M., Galeati, G. & Seren, E. (1988 a). Effect of follicle somatic cells during pig oocyte maturation on egg penetrability and male pronucleus formation. Gamete Res. 20, 117–83.CrossRefGoogle ScholarPubMed
Mattioli, M., Galeati, G., Bacci, M.L. & Seren, E. (1988 b). Follicular factors influence oocyte fertilizability by modulating the intercellular cooperation between cumulus cells and oocyte. Gamete Res. 21, 223–32.CrossRefGoogle ScholarPubMed
Mattioli, M., Galeati, G., Bacci, M.L. & Barboni, B. (1991). Changes in maturation-promoting activity in the cytoplasm of pig oocytes throughout maturation. Mol. Reprod. Dev. 30, 119–25.CrossRefGoogle ScholarPubMed
McGaughey, R.W. (1977). The culture of pig oocytes in minimal medium, and the influence of progesterone and estradiol-17β on meiotic maturation. Endocrinology 100, 39–45.CrossRefGoogle ScholarPubMed
Moor, R.M., Mattioli, M., Ding, J. & Nagai, T. (1990). Maturation in pig oocytes in vivo and in vitro. J. Reprod. Fertil, Suppl. 40, 197–210.Google ScholarPubMed
Moriya, M. & Katagiri, C. (1976). Microinjection of toad perm into oocytes undergoing maturation division. Deo. Growth Differ. 18, 349–56.CrossRefGoogle Scholar
Naito, K. & Toyoda, Y. (1991). Fluctuation of histone HI kinase activity during meiotic maturation in porcine oocytes. J. Reprod. Fertil. 93, 467–73.CrossRefGoogle Scholar
Naito, K., Daen, F.P. & Toyoda, Y. (1992). Comparison of histone H1 kinase activity during meiotic maturation between two types of porcine oocytes matured in different media in vitro. Biol. Reprod. 47, 43–7.CrossRefGoogle ScholarPubMed
Niwa, K. (1993). Effectiveness of in vitro maturation and in vitro fertilization techniques in pigs. J. Reprod. Fertil., Suppl. 48, 49–59.Google ScholarPubMed
Niwa, K. & Chang, M.C. (1975). Fertilization of rat eggs in vitro at various times before and after ovulation with special reference to fertilization of ovarian oocytes matured in vitro. J. Reprod. Fertil. 43, 435–51.CrossRefGoogle ScholarPubMed
Niwa, K., Park, C.-K. & Okuda, K. (1991). Penetration in vitro of bovine oocytes during maturation by frozen-thawed spermatozoa. J. Reprod. Fertil. 91, 329–36.CrossRefGoogle ScholarPubMed
Overstreet, J.W. & Bedford, J.M. (1974). Comparison of the penetrability of the eggs vestments in follicular oocytes, unfertilized and fertilized ova of the rabbit. Dev. Biol. 41, 185–92.CrossRefGoogle Scholar
Overstreet, J.W., Yanagimachi, R., Katz, D.F., Hayashi, K. & Hanson, F.W. (1980). Penetration of human spermatozoa into the human zona pellucida and the zona-free hamster eggs: a study of fertile donors and infertile patients. Fertil. Steril. 33, 534–42.CrossRefGoogle 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
Schmiady, H., Sperling, K., Kentenich, H. & Stauber, M. (1986). Prematurely condensed human sperm chromosomes after in vitro fertilization (IVF). Hum. Genet. 74, 441–3.CrossRefGoogle ScholarPubMed
Shibuya, E.K., Boulton, T.G., Cobb, M.H. & Ruderman, J.V. (1992). Activation of p42 MAP kinase and the release of oocytes from cell cycle arrest. EMBO J. 11, 3963–75.CrossRefGoogle ScholarPubMed
Smith, L.D. & Ecker, R.E. (1971). The interaction of steroids with Rana pipiens oocytes in the induction of maturation. Dev. Biol. 25, 232–47.CrossRefGoogle ScholarPubMed
Szollosi, D., Balakier, H., Czolowska, R. & Tarkowski, A. (1980). Ultrastructure of cell hybrids between mouse oocytes and blastomeres. J. Exp. Zool. 213, 315–25.CrossRefGoogle Scholar
Tarkowski, A.K. & Balakier, H. (1980). Nucleo-cytoplasmic interactions in cell hybrids between mouse oocytes, blastomeres and somatic cells. J. Embryol. Exp. Morphol. 55, 319–30.Google ScholarPubMed
Verlhac, M.H., de Pennart, H., Maro, B., Cobb, M.H. & Clarke, H.J. (1993). MAP kinase becomes stably activated at metaphase and is associated with microtubule-orga-nizing centers during meiotic maturation of mouse oocytes. Dev. Biol. 158, 330–40.CrossRefGoogle ScholarPubMed
Verlhac, M.H., Zubiak, J.Z., Clarke, H.J. & Maro, B. (1994). Microtubule and chromatin behaviour follow MAP kinase activity but not MPF activity during meiosis in mouse oocytes. Development 120, 1017–25.CrossRefGoogle Scholar
Wang, W.H., Niwa, K. & Okuda, K. (1991). In-vitro penetration of pig oocytes matured in culture by frozen-thawed ejaculated spermatozoa. J. Reprod. Fertil. 93, 491–6.CrossRefGoogle ScholarPubMed
Wang, W.H., Uchida, M. & Niwa, K. (1992). Effects of follicle cells on in vitro penetration of pig oocytes by cryopre-served, ejaculated spermatozoa. J. Reprod. Dev. 38, 125–31.CrossRefGoogle Scholar
Wang, W.H., Abeydeera, L.R., Okuda, K. & Niwa, K. (1994). Penetration of porcine oocytes during maturation in vitro by cryopreserved ejaculated spermatozoa. Biol. Reprod. 50, 510–15.CrossRefGoogle ScholarPubMed
Wiesel, S. & Schultz, G.A. (1981). Factors which may affect removal of protamine from sperm DNA during fertilization in the rabbit. Gamete Res. 4, 25–34.CrossRefGoogle Scholar
Yoshida, M., Bamba, K. & Kojima, Y. (1989). Effects of gonadotropins and estradiol-17β on the tuning of nuclear maturation and cumulus mass expansion in pig oocytes cultured in vitro. Jpn. J. Anim. Reprod. 35, 86–91.CrossRefGoogle Scholar
Yoshida, M., Ishigaki, K., Nagai, T., Chikyu, M. & Pursel, V.G. (1993). 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