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Stage-specific requirement of phosphate for development of rat 1-cell embryos in a chemically defined medium

Published online by Cambridge University Press:  26 September 2008

Kazuchika Miyoshi  and
Koji Niwa
Kazuchika Miyoshi
Affiliation:
Division of Animal Science and Technology, Faculty of Agriculture, Okayama University, Okayama, Japan.
Koji Niwa*
Affiliation:
Division of Animal Science and Technology, Faculty of Agriculture, Okayama University, Okayama, Japan.
*
Koji Niwa, Division of Animal Science and Technology, Faculty of Agriculture, Okayama University, 1-1-1 Tsushima-Naka, Okayama 700, Japan. Telephone: +86-251-8328. Fax: +86-254-0714.
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Summary

Rat 1-cell embryos, recovered from naturally mated females, were cultured in a chemically defined medium (mR1ECM). When examined after 24, 56, 64 and 80h of culture, embryos developed to the 2-cell (100%), 4-cell (93%), 4-cell to 8-cell (97%) and ≥8-cell (95%) stages, respectively. When 0.4 M phosphate (NaH2PO4) was added to the medium after 0, 24, 56 and 64 h of culture, percentages (0–67%) of embryos that developed to the blastocyst stage after 115h of culture were lower than that (86%) in the medium without phosphate. However, addition of phosphate after 80h of culture accelerated blastocyst formation; a significantly higher percentage (94%) of blastocysts was obtained after 110 h of culture in this medium compared with when phosphate was not added (67%). When phosphate was added to the medium after 64h, almost all (97%) 8-cell embryos developed to the blastocyst stage by 100h of culture but development of 4-cell to 7-cell embryos was inhibited (22–63%). Acceleration of blastocyst formation was caused by addition of phosphate rather than by the exchange of the medium. The stimulatory effect of phosphate on embryo development was observed at concentrations of 0.1–1.2mM. The mean numbers of cells (54.5–60.9 cells) in blastocysts examined after 115 h of culture were increased by the addition of 0.4–1.6mM phosphate at 80 h as compared with blastocysts from cultures without phosphate (46.6 cells).

Keywords

Rat1-cell embryoIn vitro developmentPhosphateChemically defined medium
Type
Article
Information
Zygote , Volume 5 , Issue 1 , February 1997 , pp. 67 - 73
Copyright
Copyright © Cambridge University Press 1997

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References

Biggers, J.D. (1987). Pioneering mammalian embryo culture. In The Mammalian Preimplantation Embryo: Regulation of Growth and Differentiation in Vitro, ed. Bavister, B.D. pp. 1–22. New York: Plenum Publishing.Google Scholar
Brison, D.R. & Leese, H.J. (1994). Blastocoel cavity formation by preimplantation rat embryos in the presence of cyanide and other inhibitors of oxidative phosphorylation. J. Reprod. Fertil.. 101, 305–9.CrossRefGoogle ScholarPubMed
Brown, J.J.G. & Whittingham, D.C. (1992). The dynamic provision of different energy substrates improves development of one-cell random-bred mouse embryos in vitro. J. Reprod. Fertil.. 95, 503–11.CrossRefGoogle ScholarPubMed
Chatot, C.L., Ziomek, C.A., Bavister, B.D., Lewis, J.L. & Torres, I. (1989). An improved culture medium supports development of random-bred 1-cell mouse embryos in vitro J. Reprod. Fertil. 86, 679–88.CrossRefGoogle ScholarPubMed
Chatot, C.L., Lewis, J.L., Torres, I. & Ziomek, C.A. (1990a). Development of 1-cell embryos from different strains of mice in CZB medium. Biol. Reprod. 42, 432–40.CrossRefGoogle ScholarPubMed
Chatot, C.L., Tasca, R.J. & Ziomek, C.A. (1990b). Glutamine uptake and utilization by preimplantation mouse embryos in CZB medium. J. Reprod. Fertil. 89, 335–46.CrossRefGoogle ScholarPubMed
Chatot, C.L., Lewis, J.L., Torres, I. & Ziomek, C.A. (1994). One-minute exposure of 4-cell mouse embryos to glucose overcomes morula block in CZB medium. Mol. Reprod. Dev. 37, 407–12.CrossRefGoogle ScholarPubMed
Crabtree, H.G. (1929). Observations on the carbohydrate metabolism of tumours. Biochem. J. 23, 536–45.CrossRefGoogle ScholarPubMed
Cross, P.C. & Brinster, R.L. (1973). The sensitivity of one-cell mouse embryos to pyruvate and lactate. Exp. Cell. Res. 77, 57–62.CrossRefGoogle Scholar
Davis, D.L. & Day, B.N. (1978). Cleavage and blastocyst formation by pig eggs in vitro J. Anim. Sci. 46, 1043–53.CrossRefGoogle ScholarPubMed
Ellington, J.E., Carney, E.W., Farrell, P.B., Simkin, M.E. & Foote, R.H. (1990) Bovine 1- to 2-cell embryo development using a simple medium in three oviduct epithelial cell coculture systems. Biol. Reprod. 43, 97–104.CrossRefGoogle Scholar
FitzGerald, L. & DiMattina, M. (1992).An improved medium for long-term culture of human embryos overcomes the in vitro developmental block and increases blastocyst formation. Fertil. Steril. 57, 641–7.CrossRefGoogle ScholarPubMed
Gardner, D.K. & Lane, M. (1993). Amino acids and ammonium regulate mouse embryo development in culture. Biol. Reprod 48, 377–85.CrossRefGoogle ScholarPubMed
Goddard, M.J. & Pratt, H.P.M (1983). Control of events during early cleavage of the mouse embryo: an analysis of the ′2-cell block′. J. Embryol. Exp. Morphol 73, 111–33.Google ScholarPubMed
Kane, M.T. & Buckley, N.J. (1977). The effects of inhibitors of energy metabolism on the growth of one-cell rabbit ova to blastocysts in vitro J. Reprod. Fertil. 49, 261–6.CrossRefGoogle ScholarPubMed
Kim, J.H., Niwa, K., Lim, J.M. & Okuda, K. (1993). Effects of phosphate, energy substrates, and amino acids on development of in vitro-matured, in vitro-fertilized bovine oocytes in a chemically defined, protein-free culture medium. Biol. Reprod. 48, 1320–5.CrossRefGoogle Scholar
King, W.A., Linares, T., Gustavsson, I. & Bane, A. (1979). A method for preparation of chromosomes from bovine zygotes and blastocysts. Vet. Sci. Comm. 3, 51–6.CrossRefGoogle Scholar
Kishi, J., Noda, Y., Narimoto, K., Umaoka, Y. & Moti, T. (1991). Block to development in cultured rat 1-cell embryos is overcome using medium HECM-1. Hum. Reprod. 6, 1445–8.CrossRefGoogle ScholarPubMed
Lane, M. & Gardner, D.K. (1994). Increase in postimplantation development of cultured mouse embryos by amino acids and induction of fetal retardation and exencephaly by ammonium ions. J. Reprod. Fertil. 102, 305–12.CrossRefGoogle ScholarPubMed
Lawitts, J.A. & Biggers, J.D. (1991). Optimization of mouse embryo culture media using simplex methods. J. Reprod. Fertil. 91, 543–56.CrossRefGoogle ScholarPubMed
Mayer, J.F. & Fritz, H.I. (1974). The culture of preimplantation rat embryos and the production of allophenic rats. J. Reprod. Fertil. 39, 1–9.CrossRefGoogle ScholarPubMed
Miyoshi, K., Funahashi, H., Okuda, K. & Niwa, K. (1994). Development of rat one-cell embryos in a chemically defined medium: effects of glucose, phosphate and osmolarity. J. Reprod. Fertil. 100, 21–6.CrossRefGoogle Scholar
Miyoshi, K., Abeydeera, L.R., Okuda, K. & Niwa, K. (1995a). Effects of osmolarity and amino acids in a chemically defined medium on development of rat one-cell embryos. J. Reprod. Fertil. 103, 27–32.CrossRefGoogle Scholar
Miyoshi, K., Tanaka, N. & Niwa, K. (1995b). Penetration in vitro of naturally ovulated rat eggs and the development of eggs in a chemically defined medium. J. Mamm. Ova Res. 12, 35–9.CrossRefGoogle Scholar
Monis, H.M. & Bavister, B.D. (1990a). Analysis of the inhibitory effect of inorganic phosphate on development of four-cell hamster embryos in vitro J. Exp. Zool. 256, 75–83.CrossRefGoogle ScholarPubMed
Monis, H.M. & Bavister, B.D. (1990b). Development of four-cell hamster embryos to the blastocyst stage in vitro and its regulation by components of the culture milieu. Reprod. Fertil. Dev. 2, 1–9.CrossRefGoogle Scholar
Petters, R.M., Johnson, B.H., Reed, M.L. & Archibong, A.E. (1990). Glucose, glutamine and inorganic phosphate in early development of the pig embryo in vitro. J. Reprod. Fertil. 89, 269–75.CrossRefGoogle ScholarPubMed
Pinyopummintr, T. & Bavister, B.D. (1991). In vitro-matured/in vitro-fertilized bovine oocytes can develop into morulae/blastocysts in chemically defined, protein-free culture media. Biol. Reprod. 45, 736–42.CrossRefGoogle ScholarPubMed
Quinn, P. (1995). Enhanced results in mouse and human embryo culture using a modified human tubal fluid medium lacking glucose and phosphate. J. Assist. Reprod. Genet. 12, 97–105.CrossRefGoogle Scholar
Quinn, P., Moinipanah, R., Steinberg, J.M. & Weathersbee, P.M. (1995). Successful human in vitro fertilization using a modified human tubal fluid medium lacking glucose and phosphate ions. Fertil. Steril. 63, 922–4.CrossRefGoogle Scholar
Schini, S.A. & Bavister, B.D. (1988). Two-cell block to development of cultured hamster embryos is caused by phosphate and glucose. Biol. Reprod. 39, 1183–92.CrossRefGoogle ScholarPubMed
Seshagiri, P.B. & Bavister, B.D. (1989a). Phosphate is required for inhibition by glucose of development of hamster 8-cell embryos in vitro Biol. Reprod. 40, 607–14.CrossRefGoogle ScholarPubMed
Seshagiri, P.B. & Bavister, B.D. (1989b). Glucose inhibits development of hamster 8-cell embryos in vitro. Biol. Reprod. 40, 599–606.CrossRefGoogle ScholarPubMed
Seshagiri, P.B. & Bavister, B.D. (1991). Glucose and phosphate inhibit respiration and oxidative metabolism in cultured hamster eight-cell embryos: evidence for the ′Crabtree effect′. Mol. Reprod. Dev. 30, 105–11.CrossRefGoogle ScholarPubMed
Slater, E.C. (1963). Uncouplers and inhibitors of oxidative phosphorylation. In Metabolic Inhibitors, vol. 11, ed. Hochster, R.M. & Quastel, J.H. pp. 503–16. New York: Academic Press.CrossRefGoogle Scholar
Slater, E.C. (1967). Application of inhibitors and uncouplers for a study of oxidative phosphorylation. Methods. Enzymol. 10, 48–57.CrossRefGoogle Scholar
Thibault, C. (1966). In vitro culture of cow egg. Ann. Biol. Anim. Biochim. Biophys. 6, 159–64.CrossRefGoogle Scholar
Thomson, J.L. (1967). Effect of inhibitors of carbohydrate metabolism on the development of preimplantation mouse embryos. Exp. Cell. Res. 46, 252–62.CrossRefGoogle ScholarPubMed
Toyoda, Y. & Chang, M.C. (1974). Fertilization of rat eggs in vitro by epididymal spermatozoa and the development of eggs following transfer. J. Reprod. Fertil. 36, 9–22.CrossRefGoogle ScholarPubMed
Whittingham, D.G. (1975). Survival of rat embryos after freezing and thawing. J. Reprod. Fertil. 43, 575–8.CrossRefGoogle ScholarPubMed
Whittingham, D.G. & Bavister, B.D. (1974). Development of hamster eggs fertilized in vitro and in vivo J. Reprod. Fertil. 38, 489–92.CrossRefGoogle Scholar
Yanagimachi, R. & Chang, M.C. (1964). In vitro fertilization of golden hamster ova. J. Exp. Zool. 156, 361–76.CrossRefGoogle ScholarPubMed
Zhang, X. & Armstrong, D.T. (1990). Presence of amino acids and insulin in a chemically defined medium improves development of 8-cell rat embryos in vitro and subsequent implantation in vivo Biol. Reprod. 42, 662–8..CrossRefGoogle Scholar