Generational Comparisons (F1 versus F3) of Vinclozolin Induced Epigenetic Transgenerational Inheritance of Sperm Differential DNA Methylation Regions (Epimutations) Using MeDIP-Seq. Rattus norvegicus

The agricultural fungicide vinclozolin exposure of a gestating female rat has previously been shown to promote transgenerational disease and epimutations in F3 generation (great-grand-offspring) animals. The current study was designed to investigate the actions of direct fetal exposure on the F1 generation rat sperm DMRs compared to the F3 transgenerational sperm DMRs. The F1 generation DMRs were found to be fewer in number and for the most part distinct from the F3 generation epimutations. Therefore, the direct exposure induced F1 generation DMRs appear to promote alterations in germ cell development that lead to the programming of the F3 generation epimutations, but are distinct between the generations. Overall design: F0 generation gestating female rats were treated with either vinclozolin or dimethyl sulfoxide (vehicle). The rats were then raised and bred to the F3 generation. Genome wide methylation levels were assessed for both the F1 and F3 generation rats. This was done by pooling three rat individual samples into a single sample pool. Three pools for each treatment in each generation were collected. The methylation for each pool was measured using MeDIP-seq on an Illumina HiSeq 2500. DOI: 10.1093/eep/dvx016