Epigenetic effects of insecticides on early differentiation of mouse embryonic stem cells

Toxicol In Vitro. 2021 Sep;75:105174. doi: 10.1016/j.tiv.2021.105174. Epub 2021 Apr 15.


Increasing evidence indicates that many insecticides produce significant epigenetic changes during embryogenesis, leading to developmental toxicities. However, the effects of insecticides on DNA methylation status during early development have not been well studied. We developed a novel nuclear phenotypic approach using mouse embryonic stem cells harboring enhanced green fluorescent protein fused with methyl CpG-binding protein to evaluate global DNA methylation changes via high-content imaging analysis. Exposure to imidacloprid, carbaryl, and o,p'-DDT increased the fluorescent intensity of granules in the nuclei, indicating global DNA methylating effects. However, DNA methylation profiling in cell-cycle-related genes, such as Cdkn2a, Dapk1, Cdh1, Mlh1, Timp3, and Rarb, decreased in imidacloprid treatments, suggesting the potential influence of DNA methylation patterns on cell differentiation. We developed a rapid method for evaluating global DNA methylation and used this approach to show that insecticides pose risks of developmental toxicity through DNA methylation.

Keywords: DNA methylation; Developmental toxicity; Mouse embryonic stem cells; Pesticides.

MeSH terms

  • Animals
  • Carbaryl / toxicity
  • Cell Cycle Proteins / genetics
  • Cell Differentiation / drug effects
  • Cells, Cultured
  • DDT / toxicity
  • DNA Methylation / drug effects*
  • DNA-Binding Proteins / genetics
  • Epigenesis, Genetic / drug effects
  • Green Fluorescent Proteins / genetics
  • High-Throughput Screening Assays / methods*
  • Insecticides / toxicity*
  • Mice
  • Mouse Embryonic Stem Cells / drug effects*
  • Mouse Embryonic Stem Cells / metabolism
  • Neonicotinoids / toxicity
  • Nitro Compounds / toxicity


  • Cell Cycle Proteins
  • DNA-Binding Proteins
  • Insecticides
  • Mbd1 protein, mouse
  • Neonicotinoids
  • Nitro Compounds
  • enhanced green fluorescent protein
  • Green Fluorescent Proteins
  • imidacloprid
  • DDT
  • Carbaryl