Mammalian Genome

, Volume 20, Issue 9, pp 664–673

Effect of IVF and laser zona dissection on DNA methylation pattern of mouse zygotes

Authors

  • Dominika D. Peters
    • Institute of Experimental GeneticsHelmholtz Zentrum München, German Research Center for Environmental Health (GmbH)
  • Konstantin Lepikhov
    • Department of Natural Sciences – Technical Faculty III FR 8.3, Biological Sciences, Institute of Genetics/EpigeneticsUniversity of Saarland
  • Karsten Rodenacker
    • Institute of Biomathematics and BiometryHelmholtz Zentrum München, German Research Center for Environmental Health (GmbH)
  • Susan Marschall
    • Institute of Experimental GeneticsHelmholtz Zentrum München, German Research Center for Environmental Health (GmbH)
  • Auke Boersma
    • Institute of Experimental GeneticsHelmholtz Zentrum München, German Research Center for Environmental Health (GmbH)
  • Peter Hutzler
    • Institute of PathologyHelmholtz Zentrum München, German Research Center for Environmental Health (GmbH)
  • Hagen Scherb
    • Institute of Biomathematics and BiometryHelmholtz Zentrum München, German Research Center for Environmental Health (GmbH)
  • Jörn Walter
    • Department of Natural Sciences – Technical Faculty III FR 8.3, Biological Sciences, Institute of Genetics/EpigeneticsUniversity of Saarland
    • Institute of Experimental GeneticsHelmholtz Zentrum München, German Research Center for Environmental Health (GmbH)
Article

DOI: 10.1007/s00335-009-9227-0

Cite this article as:
Peters, D.D., Lepikhov, K., Rodenacker, K. et al. Mamm Genome (2009) 20: 664. doi:10.1007/s00335-009-9227-0

Abstract

In vitro fertilization (IVF) and zona pellucida laser microdissection-facilitated IVF (Laser-IVF) are presently routine procedures in human assisted reproduction. The safety of these methods at the epigenetic level is not fully understood. Studies on mouse Laser-IVF embryos provide evidence that the use of Laser-IVF leads to reduced birth rate, indicating a potential harm of this technique for the embryo. Hence, the aim of this study was to examine the difference in DNA methylation pattern between IVF- and Laser-IVF-derived mouse zygotes. We examined two experimental groups of C3HeB/FeJ oocytes: (1) zona-intact and (2) laser-microdissected oocytes that were fertilized in vitro with freshly collected spermatozoa. Zygotes were fixed 5, 8, and 12 h after fertilization, and indirect immunofluorescence staining was studied using an anti-5-methylcytidine (5-MeC) antibody. The fluorescence intensities of paternal and maternal pronuclei were evaluated using the computer-assisted analysis of digital images. In addition, we performed a semiquantitative RT-PCR analysis of the presence of transcripts of three developmental marker genes, Oct4, Dab2, and Dnmt3b, in IVF- and Laser-IVF-derived blastocysts. We observed no significant differences in methylation status of the paternal genome and in the transcripts of the developmental marker genes after IVF and Laser-IVF. In conclusion, epigenetic patterns and early embryonic development are not altered by laser-assisted IVF techniques and another explanation must be sought for the poor implantation rates observed in mice.

Copyright information

© Springer Science+Business Media, LLC 2009