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Genome-wide assessment of DNA methylation alterations induced by superovulation, sexual immaturity and in vitro follicle growth in mouse blastocysts

ABSTRACT: In their attempt to fulfill the wish of having children, women who suffer from fertility issues often undergo assisted reproductive technologies (ART) such as ovarian stimulation, which has been associated with adverse health outcomes and imprinting disorders in children. However, given the crucial role of exogenous hormone stimulation in improving human infertility treatments, a more comprehensive analysis of the potential impacts on DNA methylation in embryos following ovarian stimulation is needed. Here, we provide genome-wide DNA methylation profiles of blastocysts generated after superovulation of prepubertal or adult mice, compared with blastocysts derived from non-stimulated adult mice. Additionally, we assessed the impact of the in vitro growth and maturation of oocytes on methylation in blastocysts.

ORGANISM(S): Mus musculus

PROVIDER: GSE205097 | GEO | 2023/01/19

REPOSITORIES: GEO

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Genome-wide DNA methylation assessment in mouse oocytes: effects of in vitro growth, superovulation and sexual immaturity

Project description:There is legitimate concern over epigenetic abnormalities that could be induced by procedures applied during assisted reproduction, necessitating epigenetic assessment of new methodologies. The effect of in vitro follicle culture (IFC) and superovulation on DNA methylation in the oocyte is not fully known. Here, we present the first genome-wide analysis of oocyte DNA methylation in IFC, superovulated mouse oocytes and, uniquely, we compare these data with naturally ovulated counterparts. Globally, the distinctive methylation landscape of oocytes, comprising alternating hyper- and hypomethylated domains, is preserved irrespective of the procedure. The conservation of methylation extends to the germline differential methylated regions (DMRs) of imprinted genes, necessary for their monoallelic expression in the embryo after fertilisation. However, we do detect specific, consistent and coherent differences in DNA methylation between IFC and oocytes from other groups, and between oocytes obtained after superovulation from pre-pubertal compared with sexually mature females. Several methylation differences span entire transcription units, consistent with effects on methylation downstream of transcriptional differences. Amongst these, we found alterations in Sox5, Zfp521 and Tcf4, genes for transcripton factors related to nervous system development. These observations also suggest transcriptional and epigenetic differences between oocytes from the initial and later phases of follicle recruitment. Our results provide an important reference for the use of in vitro growth and maturation, particularly from prepubertal females, in assisted reproductive technologies or fertility preservation.

2019-12-23 | GSE128656 | GEO

The use of a brief synchronization treatment after weaning, combined with superovulation, has moderate effects on the gene expression of surviving pig blastocysts

Project description:Use of combined estrus synchronization/superovulation treatments (SS) alters ovarian and en-dometrial gene expression patterns, impairing follicle and oocyte growth, fertilization, and em-bryo development. Since the impact of SS-treatments on the transcriptome of the surviving em-bryos remains undisclosed, we hereby examined gene expression changes in day 6 blastocysts that survived a brief regimen of synchronization treatment combined with superovulation. Embryos were surgically collection at day 6 after AI and transcriptome analysis performed on blas-tocyst-stage embryos with good morphology to disclose differentially expressed genes (DEGs) between groups at P-value < 0.05 and </> 1.5-fold change. Compared with the blastocysts from control (untreated) sows, the blastocysts from SS treated sows had moderate gene expression changes, with 7 pathways disrupted with a total of 10 transcripts affected, including metabolic RDH10 and SPTLC2 gene upregulation, and downregulation of oxidation related GSTK1 and GSTO1 genes. These gene expression alterations may suggest suboptimal embryo quality and could depress the embryos' response to oxidative stress, thereby impairing subsequent embryo development. These and previous findings call for avoidance of SS-treatments in embryo transfer programs.

2023-03-22 | E-MTAB-12820 | biostudies-arrayexpress

Genome-wide assessment of DNA methylation alterations induced by superovulation, sexual immaturity and in vitro follicle growth in mouse blastocysts

Project description:Genome-wide assessment of DNA methylation alterations induced by superovulation, sexual immaturity and in vitro follicle growth in mouse blastocysts

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Dynamics of mitochondrial DNA methylation in oocytes, embryos and granulosa cells

Project description:Comparison of mitochondrial DNA (mtDNA) methylation patterns in oocytes, blastocysts and ovarian granulosa cells indicates hitherto unsuspected dynamics. Oocytes and blastocysts recovered from animals subjected to ovarian stimulation and from abattoir ovaries were analyzed using bisulphite transformation of DNA followed by whole genome sequencing. The number of mtDNA copies is quite high in oocytes (200–400,000) and early embryos, resulting in very high coverage (> 3000 X) and very low p values for each of 716 cytosine-based nucleosides. Methylation was lowest in oocytes, following by blastocysts then granulosa cells and was not restricted to CpG sites but found also at CHG and CHH sites not distributed randomly. Analysis of the corresponding genomic DNA also revealed numerous methylated CHG and CHH sites, particularly in oocytes. In association with ovarian stimulation, mtDNA methylation was lower in oocytes than in blastocysts, suggesting a developmental function. RNA analysis of mitochondria encoded genes showed a significant inverse correlation between methylation and expression level for all sequences. Methylation was more extensive in mature animals than in pre-pubertal animals and the profile in granulosa cells differed strikingly from those in oocytes and blastocysts. Although preliminary, the results suggest that mtDNA methylation might play a role during gametogenesis and might be subject to regulation according to animal environment and/or age.

2019-08-26 | GSE122599 | GEO

Effect of superovulation treatment on oocyte´s DNA methylation

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Genome-wide DNA methylation assessment in mouse oocytes: effects of in vitro growth, superovulation and sexual immaturity

Project description:Genome-wide DNA methylation assessment in mouse oocytes: effects of in vitro growth, superovulation and sexual immaturity

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The trade-off hypothesis of oocyte quantity and quality in gonadotropin-stimulated ovarian regime: proteome profile of superovulated vs naturally ovulated mouse oocytes

Project description:In order to clarify if there is a tradeoff between quantity and quality of oocytes retrieved after ovarian stimulation, we compared the protein expression of mouse oocytes and embryos with vs without a background of ovarian stimulation using gonadotropins, administered according to the standard protocol in use since more than 50 years (PMID 13185277; PMID 13475597). The data we gathered contribute to a more complete understanding of embryonic gene expression, by taking into account the effect of the commonly used superovulation.

2021-12-15 | PXD021331 | Pride

The trade-off hypothesis of oocyte quantity and quality in gonadotropin-stimulated ovarian regime: proteome profile of superovulated vs naturally ovulated mouse oocytes

2021-12-14 | PXD026347 | Pride

Embryo response to different progesterone concentrations during superovulation of Holstein heifers

Project description:Greater progesterone (P4) concentrations during the follicular growth have been associated with greater quality of embryos produced by superovulated cows and heifers. However, it is yet to be determined the mechanisms by which oocyte exposure to greater P4 concentrations improves early embryo quality. The objective of this study was to evaluate the impact of different P4 concentrations during superovulation on the transcriptome profile of early bovine embryos. A total of 63 post-puberty Holstein heifers were randomly assigned into two experimental groups: High P4 (n = 31) and Low P4 (n = 32). Heifers received a pre-synchronization protocol followed by a protocol of superovulation that included the allocated P4 treatment. Embryo collection was performed 7 days post artificial insemination and embryos were evaluated for stage of embryonic development and grades of quality. Embryos graded as good/excellent quality (High P4: n = 27; Low P4: n = 27) were randomly allocated in 3 biological replicates per treatment group, balanced for stage of embryonic development. Transcriptome analysis suggested that exposure to different concentrations of P4 during superovulation promote changes in gene expression of 7 days old embryos that may be related to their developmental competence. These modifications are associated with downregulation of beta-estradiol pathway, upregulation of trophoblast-related genes and downregulation of WNT signalling pathway. Our results suggest a potential sensitivity of future embryos to follicular P4 levels but do not allow to conclude if the effect is from the oocyte, the oviduct, or the uterus response to P4.

2024-06-04 | GSE249344 | GEO

Effect of superovulation treatment on oocyte´s DNA methylation

Project description: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.

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