Temporal regulation of prenatal embryonic development by paternal imprinted loci

ABSTRACTH19andGtl2are paternal imprinted genes that are pivotal for prenatal embryonic development. Meanwhile, mouse nongrowing oocytes and sperm- or oocyte-originated haploid embryonic stem cells (haESCs) carrying bothH19andIG-DMR (differentially DNA-methylated region) deletions (DKO) that partially mimic paternal imprinting ofH19-Igf2andDlk1-Dio3can be employed as sperm replacement to efficiently support full-term embryonic development. However, howH19-DMR andIG-DMR act together to regulate embryonic development is still largely unknown. Here, using androgenetic haESC (AG-haESC)-mediated semi-cloned (SC) technology, we showed that paternalH19-DMR andIG-DMR are not essential for pre-implantation development of SC embryos generated through injection of AG-haESCs into oocytes.H19-DMR plays critical roles before 12.5 days of gestation whileIG-DMR is essential for late-gestation of SC embryos. Interestingly, we found that combined deletions ofH19andH19-DMR can further improve the efficiency of normal development of SC embryos at mid-gestation compared to DKO SC embryos. Transcriptome and histology analyses revealed thatH19andH19-DMR combined deletions rescue the placental defects. Furthermore, we showed thatH19,H19-DMR andIG-DMR deletions (TKO) give rise to better prenatal and postnatal embryonic development of SC embryos compared to DKO. Together, our results indicate the temporal regulation of paternal imprinted loci during embryonic development.