Ancestral gestational exposure to widely used neonicotinoid thiacloprid leads to global DNA methylation alterations in spermatozoa in three generations of male mice [MEDIP-seq]

Background: Neonicotinoids, a widely used class of insecticide that used is agriculture to control insect's population, are serious public concern. Objectives: In this study, we examined the capacity of a neonicotinoid, thiacloprid (thia) to induce transgenerational effects in the male mice. Methods: The pregnant outbred Swiss female mice were exposed to thia at embryonic days E6.5 to E15.5 using “0”, and “6” mg/kg/day doses. The genome-wide analysis of DNA methylation analysis showed that differentially methylated regions (DMRs) are located at the promoters and enhancers of many important developmental genes. Notably, DMRs were largely overlapping the germ cell reprograming responsive gene (GRRs) and super-enhancer elements found to be active in ES cells, testis or brain. Overall design: MEDIP-seq of spermatozoa from first, second, and third generation adults. Methylated DNA precipitation DNA-sequencing (MEDIP-seq) for methylated DNA in mouse spermatozoa was performed using 6 ug of DNA using EpiMark® Methylated DNA Enrichment Kit (Qiagen, E2600S) according to protocol provided by manufacture.