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Although the prevalence of Intracytoplasmic sperm injection (ICSI) has increased year by year, there remains concern about the safety of these procedures because of reports of the increased risk for imprinting disorders. Previous research has demonstrated that gonadotropin stimulation contributes to an increased incidence of epimutations in ICSI-derived mice. However, the epimutations in ICSI offspring after removing the effect of gonadotropin stimulation and the possibility that epimutations are reversible by developmental reprogramming has not been investigated. Our study is the first to investigate the effect of ICSI itself on methylation and exclude the effect of superovulation using the kidney tissues from the adult and old mice. We found reduced methylation and up-regulated expression of the imprinted genes,  H19 ,  Mest  and  Peg3 , in adult ICSI mice, but the above alterations observed in adult mice were not detected in old ICSI mice. At the  Snrpn  DMR, methylation status was not altered in adult ICSI-derived mice, but hypermethylation and correlated down-regulated expression of  Snrpn  were observed in old mice. In conclusion, ICSI manipulation and early embryo culture resulted in alterations of methylation in differentially methylated region of  H19 ,  Mest ,  Peg3  and  Snrpn , and the alterations were reprogrammed by developmental reprogramming.

Altered methylations of H19, Snrpn, Mest and Peg3 are reversible by developmental reprogramming in kidney tissue of ICSI-derived mice