The successful development of mammalian embryos requires both parental genomes. Nuclear transfer techniques have been adapted to generate uniparental embryos, which possess two sets of paternal or maternal genomes. These embryos fail to develop to term because of abnormal imprinted gene expression, which is not regulated by Mendelian inheritance. Uniparental embryos provide us with an important model to investigate imprinted gene function and ontogenesis. To evaluate the pre- and post-developmental ability of haploid androgenetic mouse embryos, and to analyse the expression of imprinted genes Igf2r, Asb4 and Mest in haploid androgenetic/gynogenetic blastocysts, we produced the haploid mouse embryos using the enucleation technique, examined their development at 6.5 dpc and quantified gene expression by quantitative real-time PCR. The results demonstrated that the developmental potential of haploid embryos was severely impaired and revealed that the haploid androgenones could induce the deciduas reaction, but failed to retain a live foetus at 6.5 dpc. Expression of imprinted genes Igf2r and Asb4 was unregulated in haploid androgenetic/gynogenetic blastocysts.
Increasing numbers of reports show that imprinted genes play a crucial role in fetal development, and uniparental embryos, which possess two paternally or two maternally derived pronuclei, are excellent tools for investigating the biological significance of imprinted genes. In the present study, to examine the in vitro developmental ability and expression pattern of eight imprinted genes in uniparental embryos, we produced androgenones, gynogenones, and parthenogenones using enucleation. Our data confirmed the previously observed restriction in haploid androgenetic development potential and first indicated that diploid androgenetic embryos were arrested in the 3/4-cell stage. Some imprinted genes were expressed in androgenetic, gynogenetic, and parthenogenetic blastocysts, suggesting that they were unable to maintain their imprinted expression status in uniparental embryos and that both paternal and maternal alleles are required for the specific expression of some imprinted genes.
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