MicroRNA regulation via DNA methylation during the morula to blastocyst transition in mice

Yee Ming Lee, Huei Wen Chen, Pawan Kumar Maurya, Ching Mao Su, Chii Ruey Tzeng

Research output: Contribution to journalArticlepeer-review

16 Citations (Scopus)

Abstract

Epigenetic regulation is responsible for transcriptional silencing of genes and parental imprinting. This study addresses the question whether microRNAs (miRNAs) could be affected by DNA methylation during morula-blastocyst transition. Mouse embryos were treated with/without a DNA methyltransferase inhibitor (5-aza-2′-deoxycytidine, 5-aza-dC, 10 nM-5 μM). Changes of miRNAs were analyzed by quantitative real-time (Q-PCR)-based megaplex pre-amp microRNA assays. Development from morula to blastocyst in mice was inhibited by 5-aza-dC in a dose-dependent manner (10 nM-5 μM), with half of the embryos arrested at morula stage when treated with levels of 5-aza-dC as low as 50 nM. In total, 48 down-regulated microRNAs and 17 up-regulated microRNAs (≥5-fold changes) were identified after 5-aza-dC treatment, including let-7e, mir-20a, mir-21, mir-34b, mir-128b and mir-452. Their predicted targets were selected based on software analysis, published databases and further confirmed by Q-PCR. At least eight targets, including dnmt3a, jagged 1, sp1, edg2, abcg4, numa1, tmsb10 and csf1r were confirmed. In conclusion, 5-aza-dC-modified microRNA profiles and identification of the microRNA's targets during the morula to blastocyst stage in mice provide information that helps us to explore the relationship between fertility, microRNA regulation and epigenetic intervention.

Original languageEnglish
Article numbergar072
Pages (from-to)184-193
Number of pages10
JournalMolecular Human Reproduction
Volume18
Issue number4
DOIs
Publication statusPublished - Apr 2012

Keywords

  • Blastocyst
  • DNA methylation
  • Epigenetic
  • MicroRNA
  • Morula

ASJC Scopus subject areas

  • Reproductive Medicine
  • Embryology
  • Molecular Biology
  • Genetics
  • Obstetrics and Gynaecology
  • Developmental Biology
  • Cell Biology

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