Advertisement

Histochemistry and Cell Biology

, Volume 126, Issue 2, pp 213–217 | Cite as

DNA methylation pattern in pig in vivo produced embryos

  • Josef Fulka
  • Helena Fulka
  • Tomas Slavik
  • Konosuke Okada
  • Josef FulkaJrEmail author
Original paper

Abstract

DNA methylation/demethylation pattern, determined by 5-methylcytosine (5-MeC) immunostaining, was evaluated in porcine “in vivo” produced embryos from zygote up to the blastocyst stage. In one-cell stage embryos, only the maternal pronucleus showed a positive labeling whilst the paternal pronucleus showed almost no labeling. The intensity of labeling is high until the late morula stage. Blastocysts containing less than 100 cells showed the same intensity of labeling in both the inner cell mass (ICM) nuclei and the trophectodermal (TE) cell nuclei. Interestingly, with further cell multiplication, cells of the ICM became more intensively labeled when compared to TE cells. This distinct methylation pattern is even more profound in blastocysts containing about 200–300 cells and is not caused by the difference in the cell volume of ICM and TE cells.

Keywords

Methylation DNA Embryos Pig 

Notes

Acknowledgements

Contract grant sponsors: MZe 0002701401 (JF Jr, KO), EUROCORES EuroSTELLS GACR STE/05/E004, and AVOZ50390512 (HF).

References

  1. Barton SC, Arney KL, Shi W, Niveleau A, Fundele R, Surani MA, Haaf T (2001) Genome-wide methylation patterns in normal and uniparental early mouse embryos. Hum Mol Genet 10:2983–2987PubMedCrossRefGoogle Scholar
  2. Beaujean N, Hartshorne G, Cavilla J, Taylor J, Gardner J, Wilmut I, Meehan R, Young L (2004a) Non-conservation of mammalian preimplantation methylation dynamics. Curr Biol 14:R266–R267CrossRefGoogle Scholar
  3. Beaujean N, Taylor J, Gardner J, Wilmut I, Meehan R, Young L (2004b) Effect of limited DNA methylation reprogramming in the normal sheep embryo on somatic cell nuclear transfer. Biol Reprod 71:185–193CrossRefGoogle Scholar
  4. Bourc’his D, Le Bourhis D, Patin D, Niveleau A, Comizzoli P, Renard JP, Viegas-Pequignot E (2001) Delayed and incomplete reprogramming of chromosome methylation patterns in bovine cloned embryos. Curr Biol 11:1542–1546PubMedCrossRefGoogle Scholar
  5. Chen T, Zhang YL, Jiang Y, Liu SZ, Schatten H, Chen DY, Sun QY (2004) The DNA methylation events in normal and cloned rabbit embryos. FEBS Lett 578:69–72PubMedCrossRefGoogle Scholar
  6. Dean W, Santos F, Stojkovic M, Zakhartchenko V, Walter J, Wolf E, Reik W (2001) Conservation of methylation reprogramming in mammalian development: aberrant reprogramming in cloned embryos. PNAS 98:13734–13738PubMedCrossRefGoogle Scholar
  7. Doherty AS, Bartolomei MS, Schultz RM (2002) Regulation of stage-specific nuclear translocation of Dnmt1o during preimplantation mouse development. Dev Biol 242:255–266PubMedCrossRefGoogle Scholar
  8. Fulka H, Mrazek M, Tepla O, Fulka J Jr (2004) DNA methylation pattern in human zygotes and developing embryos. Reproduction 128:703–708PubMedCrossRefGoogle Scholar
  9. Gioia L, Barboni B, Turriani M, Capacchietti G, Pistilli MG, Berardinelli P, Mattioli M (2005) The capability of reprogramming the male chromatin after fertilization is dependent on the quality of oocyte maturation. Reproduction 130:29–39PubMedCrossRefGoogle Scholar
  10. Hogan B, Beddington R, Constantini F, Lacy E (1994) Summary of mouse development. In: Hogan B, Beddington R, Constantini F, Lacy E (eds) Manipulating the mouse embryo, 2nd edn. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, pp 21–109Google Scholar
  11. Hunter RHF (1977) Physiological factors influencing ovulation, fertilization, early development and establishment of pregnancy in pigs. Br Vet J 133:461–470PubMedGoogle Scholar
  12. Li E (2002) Chromatin modification and epigenetic reprogramming in mammalian development. Nat Rev Genet 3:662–673PubMedCrossRefGoogle Scholar
  13. Mayer W, Niveleau A, Walter J, Fundele R, Haaf T (2000) Demethylation of the zygotic paternal genome. Nature 403:501–502PubMedCrossRefGoogle Scholar
  14. McLay DW, Clarke HJ (2003) Remodelling the paternal chromatin at fertilization in mammals. Reproduction 125:625–633PubMedCrossRefGoogle Scholar
  15. Morgan HD, Santos F, Green K, Dean W, Reik W (2005) Epigenetic reprogramming in mammals. Hum Mol Genet 14:R47–R58PubMedCrossRefGoogle Scholar
  16. Oswald J, Engemann S, Lane N, Mayer W, Olek A, Fundele R, Dean W, Reik W, Walter J (2000) Active demethylation of the paternal genome in the mouse zygote. Curr Biol 10:475–478PubMedCrossRefGoogle Scholar
  17. Santos F, Dean W (2004) Epigenetic reprogramming during early development in mammals. Reproduction 127:643–651PubMedCrossRefGoogle Scholar
  18. Santos F, Hendrich B, Reik W, Dean W (2002) Dynamic reprogramming of DNA methylation in the early mouse embryo. Dev Biol 241:172–182PubMedCrossRefGoogle Scholar
  19. Santos F, Zakhartchenko V, Stojkovic M, Peters A, Jenuwein T, Wolf E, Reik W, Dean W (2003) Epigenetic marking correlates with developmental potential in cloned bovine preimplantation embryos. Curr Biol 13:1116–1121PubMedCrossRefGoogle Scholar
  20. Santos F, Peters AH, Otte AP, Reik W, Dean W (2005) Dynamic chromatin modifications characterize the first cell cycle in mouse embryos. Dev Biol 280:225–236PubMedCrossRefGoogle Scholar
  21. Shi W, Haaf T (2002) Aberrant methylation patterns at the two-cell stage as an indicator of early developmental failure. Mol Reprod Dev 63:329–334PubMedCrossRefGoogle Scholar
  22. Shi W, Dirim F, Wolf E, Zakhartchenko V, Haaf T (2004) Methylation reprogramming and chromosomal aneuploidy in in vivo fertilized and cloned rabbit preimplantation embryos. Biol Reprod 71:340–347PubMedCrossRefGoogle Scholar
  23. Spinaci M, Seren E, Mattioli M (2004) Maternal chromatin remodeling during maturation and after fertilization in mouse oocytes. Mol Reprod Dev 69:215–221PubMedCrossRefGoogle Scholar
  24. Young LE, Beaujean N (2004) DNA methylation in the preimplantation embryo: the differing stories of the mouse and sheep. Anim Reprod Sci 82–83:61–78PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2006

Authors and Affiliations

  • Josef Fulka
    • 1
  • Helena Fulka
    • 2
    • 3
  • Tomas Slavik
    • 1
  • Konosuke Okada
    • 2
  • Josef FulkaJr
    • 2
    Email author
  1. 1.Center for Cell Therapy and Tissue RepairIAPG Czech Academy of SciencesLibechovCzech Republic
  2. 2.Institute of Animal ProductionPrague 114Czech Republic
  3. 3.Institute of Experimental MedicineCzech Academy of SciencesPrague 4Czech Republic

Personalised recommendations