Acrylamide (ACR), a carcinogen for rodents and humans, exists widely in the human living environment and heat-treated carbohydrate foodstuffs. Increasing evidence has demonstrated that ACR can cause chromosomal damage in somatic cells and mutagenesis. However, little is known about whether ACR can disturb genomic imprinting during spermatogenesis. We investigated the effects of ACR on methylation patterns of rat sperm genes. The results showed that after oral administration of ACR to rats for two weeks, methylation of some cytosines in the CpG islands of the differentially methylated region (DMR2) of sperm gene insulin-like growth factor II (Igf2), which is still present at the 19th day, disappeared on the 35th day. Furthermore, the extent by which ACR causes methylation defects varies in animals. Our findings indicate that mitotic spermatogonia and primary spermatocytes are sensitive to ACR-induced genomic imprinting aberration, suggesting that ACR predominantly interferes with the remodeling process in spermatogenesis.