DNA methyltransferase (Dnmt) silencing causes increased Cdx2 and Nanog levels in surviving embryos

Int J Dev Biol. 2023;67(1):1-8. doi: 10.1387/ijdb.230040oc.

Abstract

Epigenetic mechanisms are one of the essential regulators of gene expression which do not involve altering the primary nucleotide sequence. DNA methylation is considered among the most prominent epigenetic mechanisms in controlling the functions of genes related to cell differentiation, cell cycle, cell survival, autophagy, and embryo development. DNA methyl transferases (Dnmts) control DNA methylation, the levels of which are differentially altered during embryonic development, and may determine cell differentiation fate as in the case of pluripotent inner cell mass (ICM) or trophectoderm (TE). In this study, we aimed to analyze the role of Dnmt1 and Dnmt3a enzymes in ICM (using the Nanog marker) and TE (using the Cdx2 marker) differentiation, autophagy (using p62 marker), reactive oxygen species (ROS) production, and apoptosis (using TUNEL) during mouse preimplantation embryo development. Following knockdown of Dnmt1 and Dnmt3a in zygotes, expression levels of Cdx2 in the trophectoderm and Nanog in the inner cell mass were measured, as well as p62 levels, reactive oxygen species (ROS) production, and apoptosis levels after 96 hours in embryo culture. We found that knockdown of Dnmt1 or Dnmt3a significantly induced Cdx2 and Nanog expression. Similarly, p62 expression, ROS levels and apoptosis significantly increased after silencing. This study shows that Dnmt genes are highly crucial for embryonic fate determination and survival. Further studies are required to reveal the specific targets of these methylation processes related to cell differentiation, survival, autophagy, and ROS production in mouse and human preimplantation embryos.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Apoptosis
  • Blastocyst / metabolism
  • CDX2 Transcription Factor* / genetics
  • Cell Differentiation
  • DNA (Cytosine-5-)-Methyltransferase 1* / genetics
  • DNA Methyltransferase 3A* / genetics
  • Embryo, Mammalian
  • Epigenesis, Genetic
  • Female
  • Gene Silencing*
  • Mice
  • Mice, Inbred BALB C
  • Nanog Homeobox Protein* / genetics
  • Reactive Oxygen Species

Substances

  • Cdx2 protein, mouse
  • Nanog protein, mouse
  • Dnmt1 protein, mouse
  • Dnmt3a protein, mouse
  • DNA Methyltransferase 3A
  • DNA (Cytosine-5-)-Methyltransferase 1
  • Nanog Homeobox Protein
  • CDX2 Transcription Factor
  • Reactive Oxygen Species