Absence of global genomic cytosine methylation pattern erasure during medaka (Oryzias latipes) early embryo development

Ronald B. Walter, Hai Ying Li, Gabriel W. Intano, Steven Kazianis, Christi A. Walter

Research output: Contribution to journalArticle

17 Scopus citations

Abstract

Two techniques were used to analyze global genomic 5-methyl cytosine methylation at CCGG sites of medaka embryo DNA. DNA was labeled by incorporation of microinjected radiolabeled deoxynucleotide into one-cell embryos. After Hpa II or Msp I digestion the radiolabeled DNA was fractionated in agarose gels and the distribution of label quantified throughout each sample lane to detect differences in fragment distribution. Alternately isolated DNA was digested with Hpa II or Msp I and the resulting generated termini end-labeled. The end-labeled digestion products were then analyzed for fragment distribution after gel fractionation. These techniques proved to be extremely sensitive, allowing comparison of genomic DNA methylation values from as few as 640 fish cells. The data suggest that in medaka embryos the vast majority (>90%) of genomic DNA is methylated at CCGG sites. Furthermore, these data support the conclusion that the extent of methylation at these sites does not change or changes very little during embryogenesis (from 16 cells to the hatchling). These data argue against active demethylation, or loss of methylation patterns by dilution, during the developmental stages between the one cell zygote and gastrulation. From a comparative viewpoint, these data may indicate that mammals and fishes methylate and demethylate their genomes in very different manners during development.

Original languageEnglish (US)
Pages (from-to)597-607
Number of pages11
JournalComparative Biochemistry and Physiology - B Biochemistry and Molecular Biology
Volume133
Issue number4
DOIs
StatePublished - Dec 1 2002

Keywords

  • Embryogenesis
  • Fish
  • Genomic DNA
  • Medaka
  • Methylation patterns
  • Methylcytosine
  • Microinjection
  • Restriction analyses

ASJC Scopus subject areas

  • Biochemistry
  • Physiology
  • Molecular Biology

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