An Oct4-Sall4-Nanog network controls developmental progression in the pre-implantation mouse embryo

Mol Syst Biol. 2013:9:632. doi: 10.1038/msb.2012.65.

Abstract

Landmark events occur in a coordinated manner during pre-implantation development of the mammalian embryo, yet the regulatory network that orchestrates these events remains largely unknown. Here, we present the first systematic investigation of the network in pre-implantation mouse embryos using morpholino-mediated gene knockdowns of key embryonic stem cell (ESC) factors followed by detailed transcriptome analysis of pooled embryos, single embryos, and individual blastomeres. We delineated the regulons of Oct4, Sall4, and Nanog and identified a set of metabolism- and transport-related genes that were controlled by these transcription factors in embryos but not in ESCs. Strikingly, the knockdown embryos arrested at a range of developmental stages. We provided evidence that the DNA methyltransferase Dnmt3b has a role in determining the extent to which a knockdown embryo can develop. We further showed that the feed-forward loop comprising Dnmt3b, the pluripotency factors, and the miR-290-295 cluster exemplifies a network motif that buffers embryos against gene expression noise. Our findings indicate that Oct4, Sall4, and Nanog form a robust and integrated network to govern mammalian pre-implantation development.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Blastocyst / metabolism
  • Blastocyst / physiology*
  • DNA (Cytosine-5-)-Methyltransferases / genetics
  • DNA (Cytosine-5-)-Methyltransferases / metabolism
  • DNA Methyltransferase 3B
  • DNA-Binding Proteins / genetics*
  • DNA-Binding Proteins / metabolism
  • Embryo Culture Techniques
  • Embryo, Mammalian / metabolism
  • Embryonic Development
  • Embryonic Stem Cells / physiology*
  • Female
  • Gene Expression Profiling
  • Gene Expression Regulation, Developmental
  • Gene Knockdown Techniques
  • Gene Regulatory Networks*
  • Homeodomain Proteins / genetics*
  • Homeodomain Proteins / metabolism
  • Male
  • Mice
  • Mice, Inbred C57BL
  • Mice, Inbred DBA
  • MicroRNAs / genetics
  • Nanog Homeobox Protein
  • Octamer Transcription Factor-3 / genetics*
  • Octamer Transcription Factor-3 / metabolism
  • Oligonucleotide Array Sequence Analysis
  • Transcription Factors / genetics*
  • Transcription Factors / metabolism

Substances

  • DNA-Binding Proteins
  • Homeodomain Proteins
  • MIRN290 microRNA, mouse
  • MIRN295 microRNA, mouse
  • MicroRNAs
  • Nanog Homeobox Protein
  • Nanog protein, mouse
  • Octamer Transcription Factor-3
  • Sall4 protein, mouse
  • Transcription Factors
  • DNA (Cytosine-5-)-Methyltransferases