Czech J. Anim. Sci., 2012, 57(2):83-94 | DOI: 10.17221/5135-CJAS

Dynamics of epigenetic remodeling in interspecies porcine zygotes

I. Barnetova1,2, I. Vackova1,2, P. Firla1
1 Institute of Animal Science, Prague-Uhříněves, Czech Republic
2 Center for Cell Therapy and Tissue Repair, Prague, Czech Republic

The process of active paternal chromatin demethylation after fertilization in the pig is not fully understood and very inconsistent data have been published by different research groups. We have applied the interspecies intracytoplasmic sperm injection (iICSI) to evaluate remodeling capabilities of porcine oocytes in more details. We injected mouse frozen-thawed sperm heads into porcine in vitro matured or ovulated oocytes, respectively. Embryos produced by intracytoplasmic sperm injection (ICSI) of boar spermatozoa into porcine ovulated oocytes (intraspecies) served as controls. Zygotes with 2-pronuclei were labeled with antibodies against certain epigenetic modifications (5-methylcytosine, 5-MeC; heterochromatin protein 1, HP1; trimethylation of H3/K4, H3/K4-me3; and dimethylation of H3/K9, H3/K9-me2). The labeling patterns were not different between zygotes produced from in vitro matured and ovulated oocytes. Both pronuclei were symmetrically labeled with 5-MeC, HP1, and H3/K4-me3 antibodies. Asymmetrical labeling was observed only with H3/K9-me2 antibody. The labeling of interspecies zygotes was similar to that of intraspecies zygotes. Moreover, the DNA demethylation was observed neither in control zygotes (intraspecies). The only difference observed between zygotes produced from in vitro matured and ovulated oocytes was in their ability to be activated. Intraspecies zygotes produced from ovulated oocytes were able to form the paternal pronucleus without additional activation; the zygotes produced from in vitro matured oocytes formed the paternal pronucleus only after additional activation with electric pulses. Our results show that the remodeling abilities of in vitro matured and ovulated oocytes are essentially similar. Moreover, it seems that reasons of inconsistent data reporting the active demethylation in the pig are more complicated and they are not associated exclusively with the oocyte quality.

Keywords: pig; methylation; iICSI; zygote

Published: February 29, 2012  Show citation

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Barnetova I, Vackova I, Firla P. Dynamics of epigenetic remodeling in interspecies porcine zygotes. Czech J. Anim. Sci. 2012;57(2):83-94. doi: 10.17221/5135-CJAS.
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    References

    1. Abdalla H., Hirabayashi M., Hochi S. (2009): Demethylation dynamics of the paternal genome in pronuclearstage bovine zygotes produced by in vitro fertilization and ooplasmic injection of freeze-thawed or freezedried spermatozoa. Journal of Reproduction and Development, 55, 433-439. Go to original source... Go to PubMed...
    2. Adenot P.G., Mercier Y., Renard J.P., Thompson E.M. (1997): Differential H4 acetylation of paternal and maternal chromatin precedes DNA replication and differential transcriptional activity in pronuclei of 1-cell mouse embryos. Development, 124, 4615-4625. Go to original source... Go to PubMed...
    3. Araki Y., Yoshizawa M., Araky Y. (2005): A novel method for chromosome analysis of human sperm using enucleated mouse oocytes. Human Reproduction, 20, 1244-1247. Go to original source... Go to PubMed...
    4. Barnetova I., Okada K. (2010): Genome reprogramming during the first cell cycle in in vitro produced porcine embryos. Czech Journal of Animal Science, 55, 49-57. Go to original source...
    5. Barnetova I., Fulka H., Fulka Jr., J. (2010): Epigenetic characteristics of paternal chromatin in interspecies zygotes. Journal of Reproduction and Development, 56, 601-606. Go to original source... Go to PubMed...
    6. Beaujean N., Hartshorne G., Cavilla J., Taylor J., Gardner J., Wilmut I., Meehan R., Young L. (2004a): Non-conservation of mammalian preimplantation methylation dynamics. Current Biology, 14, R266-R267. Go to original source... Go to PubMed...
    7. Beaujean N., Taylor J.E., McGarry M., Gardner J.O., Wilmut I., Loi P., Ptak G., Galli C., Lazzari G., Bird A., Young L.E., Meehan R.R. (2004b): The effect of interspecific oocytes on demethylation of sperm DNA. Proceedings of the National Academy of Sciences, 101, 7636-7640. Go to original source... Go to PubMed...
    8. 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. Proceedings of the National Academy of Sciences, 98, 13734-13738. Go to original source... Go to PubMed...
    9. Deshmukh R.S., Ostrup O., Ostrup E., Vejlsted M., Niemann H., Lucas-Hahn A., Petersen B., Li J., Callesen H., Hyttel P. (2011): DNA methylation in porcine preimplantation embryos developed in vivo and produced by in vitro fertilization, parthenogenetic activation, and somatic cell nuclear transfer. Epigenetics, 6, 177-187. Go to original source... Go to PubMed...
    10. Fulka H., Mrazek M., Tepla O., Fulka Jr., J. (2004): DNA methylation pattern in human zygotes and developing embryos. Reproduction, 128, 703-708. Go to original source... Go to PubMed...
    11. Fulka H., Fulka J. (2006): No differences in the DNA methylation pattern in mouse zygotes produced in vivo, in vitro, or by intracytoplasmic sperm injection. Fertility and Sterility, 86, 1534-1536. Go to original source... Go to PubMed...
    12. Fulka J., Fulka H., Slavik T., Okada K. (2006): DNA methylation pattern in pig in vivo produced embryos. Histochemistry and Cell Biology, 126, 213-217. Go to original source... Go to PubMed...
    13. Fulka H., Barnetova I., Mosko T., Fulka Jr., J. (2008): Epigenetic analysis of human spermatozoa after their injection into ovulated mouse oocytes. Human Reproduction, 23, 627-634. Go to original source... Go to PubMed...
    14. Gioia L., Barboni B., Turriani M., Capacchietti G., Pistilli M.G., Berardinelli P., Mattioli M. (2005): The capability of reprogramming the male chromatin after fertilization is dependent on the quality of oocyte maturation. Reproduction, 130, 29-39. Go to original source... Go to PubMed...
    15. Jeong Y.S., Yeo S., Park J.S., Koo D.B., Chang W.K., Lee K.K., Kang Y.K. (2007a): DNA methylation state is preserved in the sperm-derived pronucleus of the pig zygote. International Journal of Developmental Biology, 51, 707-714. Go to original source... Go to PubMed...
    16. Jeong Y.S., Yeo S., Park J.S., Lee K.K., Kang Y.K. (2007b): Gradual development of a genome-wide H3-K9 trimethylation pattern in paternally derived pig pronucleus. Developmental Dynamics, 236, 1509-1516. Go to original source... Go to PubMed...
    17. Kimura Y., Yanagimachi R. (1995): Intracytoplasmic sperm injection in the mouse. Biology of Reproduction, 52, 709-720. Go to original source... Go to PubMed...
    18. Lepikhov K., Zakhartchenko V., Hao R., Yang F., Wrenzycki C., Niemann H., Wolf E., Walter J. (2008): Evidence for conserved DNA and histone H3 methylation reprogramming in mouse, bovine and rabbit zygotes. Epigenetics & Chromatin, 1, 8. Go to original source... Go to PubMed...
    19. Mayer W., Niveleau A., Walter J., Fundele R., Haaf T. (2000): Demethylation of the zygotic paternal genome. Nature, 403, 501-502. Go to original source... Go to PubMed...
    20. 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. Current Biology, 10, 475-478 Go to original source... Go to PubMed...
    21. Park J.S., Jeong Y.S., Shin S.T., Lee K.K., Kang Y.K. (2007): Dynamic DNA methylation reprogramming: active demethylation and immediate remethylation in the male pronucleus of bovine zygotes. Developmental Dynamics, 236, 2523-2533. Go to original source... Go to PubMed...
    22. Santos F., Peters A.H., Otte A.P., Reik W., Dean W. (2005): Dynamic chromatin modifications characterise the first cell cycle in mouse embryos. Developmental Biology, 280, 225-236. Go to original source... Go to PubMed...
    23. Schatten G., Simerly C., Palmer D.K., Margolis R.L., Maul G., Andrews B.S., Schatten H. (1988): Kinetochore appearance during meiosis, fertilization and mitosis in mouse oocytes and zygotes. Chromosoma, 96, 341-352. Go to original source... Go to PubMed...
    24. Wright S.J., Longo F.J. (1988): Sperm nuclear enlargement in fertilized hamster eggs is related to meiotic maturation of the maternal chromatin. Journal of Experimental Zoology, 247, 155-165. Go to original source... Go to PubMed...
    25. Yang J., Yang S., Beaujean N., Niu Y., He X., Xie Y., Tang X., Wang L., Zhou Q., Ji W. (2007): Epigenetic marks in cloned rhesus monkey embryos: Comparison with counterparts produced in vitro. Biological Reproduction, 76, 36-42. Go to original source... Go to PubMed...
    26. Zaitseva I., Zaitsev S., Alenina N., Bader M., Krivokharchenko A. (2007): Dynamics of DNA-demethylation in early mouse and rat embryos developed in vivo and in vitro. Molecular Reproduction & Development, 74, 1255-1261. Go to original source... Go to PubMed...

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