| IVF(in vitro fertilization) are increasingly used to help infertile couples to obtain offspring. However, the security issues can not be ignored. Some epidemiological studies and animal models have demonstrated that IVF could lead to a series of health problems, including embryonic loss, preterm birth, perinatal mortality, lower birth weight and increased risk of childhood and adulthood diseases. During the past several years, high-throughput analyses, such as RNA-seq and microarrays, have enabled the transcriptome of IVF embryos to be profiled in the mouse, bovine, and sheep. MiRNA genomic, a new field research focuses on aspects of human disease, while in the rat, sheep, cattle, there are many reports. Using tandem mass spectrometry (MS/MS) based methods, the proteome of IVF embryos have also been characterized in the mouse, bovine, and porcine. In these studies, numerous differentially expressed genes, miRNA or proteins were reported and may potentially be the source of aberrant intrauterine growth, abnormal postnatal development and increased disease risk in offspring. However, the majority of studies in this field have focused on the embryos or conceptus, with only a limited number of investigations conducted on the placenta especially traced back to the early formation of the placenta, which known to be critical for normal fetal development and growth.Using mice as model, we collected extra-embryonic/placenta tissues at E7.5and El0.5, and study the effect of in vitro fertilization and culture on the epigenetic modification and gene expression patterns through High-throughput sequencing(Transcriptome, miRNA, proteome). At the same time, bioinformatics was used to explore the abnormal molecular mechanisms of IVF tissues. As well, we also recorded the phenotypes of in vivo and in vitro produced embryos during sampling process. Our results showed that significant difference can be seen at E7.5. Comparing with IVO (in vivo fertilization and development) group, the weight of embryo IVF group is lower and indicate stunting, degradation. Abnormal angiogenesis was appeared at E10.5and could be sustained to E19. In addition, in vitro produced fetus showed a lower body weight and an increased placental weight at El9, implying that the development of both embryonic and extra-embryonic tissues was compromised in vitro produced placenta.From the epigenetic modifications, Our study reveals the molecular mechanisms of in IVO and in IVF extra-embryonic tissues/placenta gene expression regulation. We analyzed data of E7.5and E10.5using transcriptome, proteome and miRNA. The results showed that numerous transcriptome, proteome differentially expressed genes (DEG) or proteins were found to be involved in the terms of metabolism, angiogenesis and hematopoiesis, transport, cytoskeleton. It was also found numerous miRNA were regulated pathways above. And the same time we achieved many important miRNA regulation patterns in IVF groups. Meanwhile, we found global hypomethylation expression patterns in IVF groups. This investigation provides the first propose in IVF abnormalities from post-translational dysregulation.Our study, for the first time, using high-throughput approaches at three levels, profiled the different expression patterns between in IVO and in IVF produced extra-embryonic tissues/placenta at E7.5and E10.5. This study provides systematic exposition of the underlying mechanisms responsible for the placentas and fetal aberrant development, and a lot of candidate genes, epigenetic modification patterns. These founds would provide a reference for further optimizing in vitro culture system. |
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