Skip to main content Accessibility help
×
×
Home

Deficiencies in extrusion of the second polar body due to high calcium concentrations during in vitro fertilization in inbred C3H/He mice

  • Yuki Ohta (a1) (a2) (a3) (a4), Yoshikazu Nagao (a3), Naojiro Minami (a5), Satoshi Tsukamoto (a2) and Seiji Kito (a6) (a2)...

Summary

Successful in vitro fertilization (IVF) of all inbred strains of laboratory mice has not yet been accomplished. We have previously shown that a high calcium concentration improved IVF in various inbred mice. However, we also found that in cumulus-free ova of C3H/He mice such IVF conditions significantly increased the deficiency of extrusion of the second polar body (PBII) in a dose-dependent manner (2% at 1.71 mM and 29% at 6.84 mM, P < 0.05) and that PBII extrusion was affected by high calcium levels at 2–3 h post-insemination. While developmental competence of ova without PBII extrusion to blastocysts after 96 h culture was not affected, a significant reduction in the nuclear number of the inner cell mass was observed in blastocyst fertilized under high calcium condition. We also examined how high calcium concentration during IVF affects PBII extrusion in C3H/He mice. Cumulus cells cultured under high calcium conditions showed a significantly alleviated deficient PBII extrusion. This phenomenon is likely to be specific to C3H/He ova because deficient PBII extrusion in reciprocal fertilization between C3H and BDF1 gametes was observed only in C3H/He ova. Sperm factor(s) was still involved in deficient PBII extrusion due to high calcium concentrations, as this phenomenon was not observed in ova activated by ethanol. The cytoskeletal organization of ova without PBII extrusion showed disturbed spindle rotation, incomplete formation of contractile ring and disturbed localization of actin, suggesting that high calcium levels affect the anchoring machinery of the meiotic spindle. These results indicate that in C3H/He mice high calcium levels induce abnormal fertilization, i.e. deficient PBII extrusion by affecting the cytoskeletal organization, resulting in disturbed cytokinesis during the second meiotic division. Thus, use of high calcium media for IVF should be avoided for this strain.

Copyright

Corresponding author

All correspondence to: S. Kito. Department of Technical Support and Development, National Institute of Radiological Sciences, 4–9-1 Anagawa Inage-ku, Chiba, 263–8555, Japan. Tel: +81 43 206 3059. Fax: +81 43 206 4093. E-mail: sk126@nirs.go.jp

References

Hide All
Altman, P.L. & Katz, D.D. (1979). Part 1 mouse and rat. I. Mice. In Inbred and Genetically Defined Strains of Laboratory Animals, pp. 9229. Federation of American Society for Experimental Biology, Bethesda, MD, USA.
Azoury, J., Verlhac, M.H. & Dumont, J. (2009) Actin filaments: key players in the control of asymmetric divisions in mouse oocytes. Biol. Cell 101, 6976.
Biggers, J., Lynda, K. McGinnis, L. & Raffin, M (2000). Amino acids and preimplantation development of the mouse in protein-free potassium simplex optimized medium. Biol. Reprod. 63, 281–93.
Brockmann, C., Huarte, J., Dugina, V., Challet, L., Rey, E., Conne, B., Swetloff, A., Nef, S., Chaponnier, C. & Vassalli, J.D. (2011). Beta- and gamma-cytoplasmic actins are required for meiosis in mouse oocytes. Biol. Reprod. 85, 1025–39.
Brunet, S. & Verlhac, M.H. (2011). Positioning to get out of meiosis: the asymmetry of division. Hum. Reprod. Update 17, 6875.
Byers, S.L., Payson, S.J. & Taft, R.A. (2006). Performance of ten inbred mouse strains following assisted reproductive technologies (ARTs). Theriogenology 65, 1716–26.
Chaigne, A., Verlhac, M.H. & Terret, M.E. (2012). Spindle positioning in mammalian oocytes. Exp. Cell Res. 318, 1442–7.
Critser, J.K. & Mobraaten, L.E. (2000). Cryopreservation of murine spermatozoa. ILAR J. 41, 197206.
Cuthbertson, K.S.R, Whittingham, D.G. & Cobbold, P.H. (1981) Free Ca2+ increases in exponential phases during mouse oocyte activation. Nature 294, 754–7.
Deng, M.Q. & Fan, B.Q. (1996). Intracellular Ca2+ during fertilization and artificial activation in mouse oocytes. Zhongguo Yao Li Xue Bao 17, 357–60.
Ducibella, T. & Fissore, R. (2008). The roles of Ca2+, downstream protein kinases, and oscillatory signaling in regulating fertilization and the activation of development. Dev. Biol. 315, 257–79.
Ducibella, T., Schultz, R.M. & Ozil, J.P. (2006) Role of calcium signals in early development. Semin Cell Dev Biol. 17, 324–32.
Fraser, L.R. (1987). Minimum and maximum extracellular Ca2+ requirements during mouse sperm capacitation and fertilization in vitro . J. Reprod. Fertil. 81, 7789.
Fraser, L.R. (1993). In vitro capacitation and fertilization. Method Enzymol. 225, 239–53
Gaunt, S.J. (1983). Spreading of a sperm surface antigen within the plasma membrane of the egg after fertilization in the rat. J. Embryol. Exp. Morphol. 75, 259–70.
Gundersen, G.G., Medill, L. & Shapiro, B.M. (1986). Sperm surface proteins are incorporated into the egg membrane and cytoplasm after fertilization. Dev. Biol. 113, 207–17.
Hino, T., Oda, K., Nakamura, K., Tateno, H., Toyoda, Y. & Yokoyama, M. (2011). Accelerated modification of the zona pellucida is the primary cause of decreased fertilizability of oocytes in the 129 inbred mouse strain. Zygote 19, 315–22.
Ho, Y., Wigglesworth, K., Eppig, J.J. & Schultz, R.M. (1995). Preimplantation development of mice embryos in KSOM: augmentation by amino acids and analysis of gene expression. Mol. Reprod. Dev. 41, 232–8.
Huneau, D. & Crozet, N. (1989). In vitro fertilization in the sheep: effect of elevated calcium concentration at insemination. Gamete Res. 23, 119–25.
Itagaki, Y. & Toyoda, Y. (1992). Effects of prolonged sperm preincubation and elevated calcium concentration on fertilization of cumulus-free mouse eggs in vitro . J. Reprod. Dev. 38, 219–24.
Johnson, J., Bierle, B.M., Gallicano, G.I. & Capco, D.G. (1998). Calcium/calmodulin-dependent protein kinase II and calmodulin: regulators of the meiotic spindle in mouse eggs. Dev. Biol. 204, 464–77.
Kaplan, R. & Kraicer, P.F. (1978). Effect of elevated calcium concentration on fertilization of rat oocyte in vitro . Gamete Res. 1, 281–5.
Kaufman, M.H. (1982). The chromosome complement of single-pronuclear haploid mouse embryos following activation by ethanol treatment. J. Embryol. Exp. Morphol. 71, 139–54.
Kim, E., Yamashita, M., Kimura, M., Honda, A., Kashiwabara, S., & Baba, T. (2008). Sperm penetration through cumulus mass and zona pellucida. Int. J. Dev. Biol. 52, 677–82.
Kito, S., Hayao, T., Noguchi-Kawasaki, Y., Ohta, Y., Hideki, U. & Tateno, S. 2004. Improved in vitro fertilization and development by use of modified human tubal fluid and applicability of pronucleate embryos for cryopreservation by rapid freezing in inbred mice. Comp. Med. 54, 564–70.
Kito, S. & Ohta, Y. (2005). Medium effects on capacitation and sperm penetration through the zona pellucida in inbred BALB/c spermatozoa. Zygote 13, 145–53.
Kito, S. & Ohta, Y. (2008a). In vitro fertilization in inbred BALB/c mice I: isotonic osmolarity and increased calcium-enhanced sperm penetration through the zona pellucida and male pronuclear formation. Zygote 16, 249–57.
Kito, S. & Ohta, Y. (2008b). In vitro fertilization in inbred BALB/c mice II: effects of lactate, osmolarity and calcium on in vitro capacitation. Zygote 16, 259–70.
Krauchunas, A.R. & Wolfner, M.F. (2013). Molecular changes during egg activation. Curr. Top. Dev. Biol. 102, 267–92.
Lawitts, J.A. & Biggers, J.D. (1993). Culture of preimplantation embryos. Methods Enzymol. 225, 153–64.
Leese, H.J. & Barton, A.M. (1985). Production of pyruvate by isolated mouse cumulus cells. J. Exp. Zool. 234, 231–6.
Marangos, P., Fitzharris, G. & Carroll, J. (2003) Ca2+ oscillations at fertilization in mammals are regulated by the formation of pronuclei. Development 130, 1461–72.
Markoulaki, S., Matson, S., Abbott, A.L. & Ducibella, T. (2003). Oscillatory CaMKII activity in mouse egg activation. Dev. Biol. 258, 464–74.
Miao, Y.L., Stein, P., Jefferson, W.N., Padilla-Banks, E. & Williams, C.J. (2012). Calcium influx-mediated signaling is required for complete mouse egg activation. Proc. Natl. Acad. Sci. USA 109, 4169–74.
Miyamoto, H. & Ishibashi, T. (1975). The role of calcium ions in fertilization of mouse and rat eggs in vitro . J. Reprod. Fertil. 45, 523–6.
Nagasawa, H., Miyamoto, M. and Fujimoto, M. (1973). Reproductivity in inbred strains of mice and project for their efficient production. Exp. Animals (Japan) 22, 119–26.
Oh, S.H., Miyoshi, K. & Funahashi, H. (1998) Rat oocytes fertilized in modified rat 1-cell embryo culture medium containing a high sodium chloride concentration and bovine serum albumin maintain developmental ability to the blastocyst stage. Biol. Reprod. 59, 884–9.
Ozil, J.P., Banrezes, B., Tóth, S., Pan, H. & Schultz, R.M. (2006). Ca2+ oscillatory pattern in fertilized mouse eggs affects gene expression and development to term. Dev. Biol. 300, 534–44.
Ozil, J.P., Markoulaki, S., Tóth, S., Matson, S., Banrezes, B., Knott, J.G., Schultz, R.M., Huneau, D. & Ducibella, T. (2005). Egg activation events are regulated by the duration of a sustained [Ca2+]cyt signal in the mouse. Dev. Biol. 282, 3954.
Quinn, P., Warnes, G.M., Kerin, J.F. & Kirby, C. (1985) Culture factors affecting the success rate of in vitro fertilization and embryo transfer. Ann. N. Y. Acad. Sci. 442, 195204.
Quinn, P. & Whittingham, D.G. (1982) Effect of fatty acids on fertilization and development of mouse embryos in vitro . J. Androl. 3, 440444
Rogers, N.T., Halet, G., Piao, Y., Carroll, J., Ko, M.S. & Swann, K. (2006). The absence of a Ca2+ signal during mouse egg activation can affect parthenogenetic preimplantation development, gene expression patterns, and blastocyst quality. Reproduction 132, 4557.
Saunders, C.M., Larman, M.G., Parrington, J., Cox, L.J., Royse, J., Blayney, L.M., Swann, K. & Lai, F.A. (2002). PLC zeta: a sperm-specific trigger of Ca2+ oscillations in eggs and embryo development. Development 129, 3533–44.
Simon, M.M., Greenaway, S., White, J.K., Fuchs, H., Gailus-Durner, V., Wells, S., Sorg, T., Wong, K., Bedu, E., Cartwright, E.J., Dacquin, R., Djebali, S., Estabel, J., Graw, J., Ingham, N.J., Jackson, I.J., Lengeling, A., Mandillo, S., Marvel, J., Meziane, H., Preitner, F., Puk, O., Roux, M., Adams, D.J., Atkins, S., Ayadi, A., Becker, L., Blake, A., Brooker, D., Cater, H., Champy, M.F., Combe, R., Danecek, P., di Fenza, A., Gates, H., Gerdin, A.K., Golini, E., Hancock, J.M., Hans, W., Hölter, S.M., Hough, T., Jurdic, P., Keane, T.M., Morgan, H., Müller, W., Neff, F., Nicholson, G., Pasche, B., Roberson, L.A., Rozman, J., Sanderson, M., Santos, L., Selloum, M., Shannon, C., Southwell, A., Tocchini-Valentini, G.P., Vancollie, V.E., Westerberg, H., Wurst, W., Zi, M., Yalcin, B., Ramirez-Solis, R., Steel, K.P., Mallon, A.M., de Angelis, M.H., Herault, Y. & Brown, S.D. (2013). A comparative phenotypic and genomic analysis of C57BL/6J and C57BL/6N mouse strains. Genome Biol. 14, R82.
Simpson, E.M., Linder, C.C., Sargent, E.E., Davisson, M.T., Mobraaten, L.E. & Sharp, J.J. (1997). Genetic variation among 129 substrains and its importance for targeted mutagenesis in mice. Nat. Genet. 16, 1927.
Sun, Q.Y. & Schatten, H. (2006). Regulation of dynamic events by microfilaments during oocyte maturation and fertilization. Reproduction 131, 193205.
Sztein, J.M., Farley, J.S. & Mobraaten, L.E. (2000). In vitro fertilization with cryopreserved inbred mouse sperm. Biol. Reprod. 63, 1774–80.
Takagi, N., Wake, N. & Sasaki, M. (1978). Cytologic evidence for preferential inactivation of the paternally derived X chromosome in XX mouse blastocysts. Cytogenet. Cell Genet. 20, 240–8.
Tatone, C., Delle Monache, S., Iorio, R., Caserta, D., Di Cola, M. & Colonna, R. (2002) Possible role for Ca2+ calmodulin-dependent protein kinase II as an effector of the fertilization Ca2+ signal in mouse oocyte activation. Mol. Hum. Reprod. 8, 750–7.
Thornton, C.E., Brown, S.D. & Glenister, P.H. (1999). Large numbers of mice established by in vitro fertilization with cryopreserved spermatozoa: implications and applications for genetic resource banks, mutagenesis screens, and mouse backcrosses. Mamm. Genome 10, 987–92.
Threadgill, D.W., Yee, D., Matin, A., Nadeau, J.H. & Magnuson, T. (1997) Genealogy of the 129 inbred strains: 129/SvJ is a contaminated inbred strain. Mamm. Genome 8, 390–3.
Toyoda, Y., Yokoyama, M. & Hoshi, T. (1971). Studies on fertilization of mouse eggs in vitro. I. In vitro fertilization of eggs by fresh epididymal sperm. Jpn J. Anim. Reprod. 16, 147–51.
Tóth, S., Huneau, D., Banrezes, B. & Ozil, J.P. (2006). Egg activation is the result of calcium signal summation in the mice. Reproduction 131, 2734.
Vasudevan, K., Raber, J. & Sztein, J. (2010). Fertility comparison between wild type and transgenic mice by in vitro fertilization. Transgenic Res. 19, 587–94.
Wakai, T., Vanderheyden, V. & Fissore, R.A. (2011). Ca2+ signaling during mammalian fertilization: requirements, players, and adaptations. Cold Spring Harb. Perspect. Biol. 3, pii: a006767.
Wang, C. & Machaty, Z. (2013). Calcium influx in mammalian eggs. Reproduction 145, R97R105.
Yanagimachi, R. (1994). Mammalian fertilization. In The Physiology of Reproduction, 2nd edn (eds Knobil, E. & Neill, J.D.), pp. 189317. New York: Raven Press.
Zar, J.H. 1996. Biostatistical Analysis, 3rd edn, pp. 277305. Englewood Cliffs, NJ: Prentice-Hall.
Recommend this journal

Email your librarian or administrator to recommend adding this journal to your organisation's collection.

Zygote
  • ISSN: 0967-1994
  • EISSN: 1469-8730
  • URL: /core/journals/zygote
Please enter your name
Please enter a valid email address
Who would you like to send this to? *
×

Keywords

Metrics

Altmetric attention score

Full text views

Total number of HTML views: 0
Total number of PDF views: 0 *
Loading metrics...

Abstract views

Total abstract views: 0 *
Loading metrics...

* Views captured on Cambridge Core between <date>. This data will be updated every 24 hours.

Usage data cannot currently be displayed