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Effect of superovulation treatment on oocyte´s DNA methylation

ABSTRACT: Controlled ovarian stimulation is a necessary step in some assisted reproductive procedures allowing the collection of female gametes at a precise time and at a suitable amount needed for subsequent fertilization steps. However, studies in animals have shown many consequences of hormonal treatments over the oocyte. Several of these studies used comparisons between oocytes from different donors, which is known that may influence the response differently. In this work we propose the use of the bovine model where each animal served as its own control. DNA methylation profile of oocytes from pre-ovulatory unstimulated follicles (NS) was compared to oocytes from stimulated follicles (S), after single-cell whole genome bisulphite sequencing. Results show that global percentage of methylation was similar between groups but percentage of methylation was lower for non-stimulated oocytes in the imprinted genes APEG3, MEG3 and MEG9, and higher in TSSC4 when compared to stimulated oocytes. Differences were also found in CpGi of imprinted genes: higher methylation was found among NS oocytes in MEST (PEG1)), IGF2R, GNAS (SCG6), KvDMR1 ICR UMD and IGF2. In another region around IGF2 the methylation percentage was lower for NS oocytes when compared to S oocytes. Data drawn from this study might help to understand the molecular reasons of the appearance of certain syndromes in assisted reproductive technologies-derived offspring.

ORGANISM(S): Bos taurus

PROVIDER: GSE166130 | GEO | 2023/01/11

REPOSITORIES: GEO

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Project description:Assisted reproductive technology (ART) is an important part of reproductive medicine that comprises superovulation, in vitro fertilization (IVF), in vitro culture of embryos, cryopreservation and embryo transfer. Because it occurs in the sensitive period of gametogenesis and early embryonic development, its possible health effects on the offspring have received widespread attention in recent years. Relevant studies, though, are limited to postnatal follow-up and blood sample analysis. In this study, a mouse model was used to explore the effect of ART on gene expression in the organs of the off-spring using next-generation sequencing. The results showed that it caused abnormal expression in 1060 and 179 genes in the heart and spleen, respectively. Differentially expressed genes (DEGs) in the heart are mainly enriched in RNA synthesis and processing, and cardiovascular system development is also enriched. STRING analysis identified Ccl2, Ptgs2, Rock1, MAPK14, Agt, Wnt5a as core interaction factors. DEGs in the spleen are significantly enriched in anti-infection and im-mune responses and in the hematopoiesis-related pathway "hematopoietic cell lineage", which includes the core factors Fos, Jun and Il1r2. Then, we explored the effects of ART on epigenetic modifiers and imprinted genes. The results showed that 42 heart genes and 5 spleen genes were abnormally expressed, most of which were related to histone modification and chromatin remodeling. Imprinted genes are very important for normal fetal and individual growth and development. It was found that the expression of imprinted genes Dhcr7, Igf2, Mest and Smoc1 decreased in the heart of ART offspring, and the DNA methylation levels of Igf2 and Mest imprinting control regions (ICRs) increased abnormally. Therefore, we con-cluded that, in the mouse model, ART can interfere with the gene expression pattern in the heart and spleen of the adult offspring, causing potentially adverse effects on organ function. The results of this study can provide a reference for evaluating the safety of human ART and helping improve the clinical system.

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Effect of superovulation treatment on oocyte´s DNA methylation

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