Microarray analysis of bovine oocyte from distinct follicle sizes reveals gradual transcripts accumulation
Ontology highlight
ABSTRACT:
ORGANISM(S): Bos taurus
PROVIDER: GSE48283 | GEO | 2015/11/15
SECONDARY ACCESSION(S): PRJNA209868
REPOSITORIES: GEO
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Project description: Not available
| S-EPMC12490830 | biostudies-literature
Project description:Vitrification is increasingly used to cryopreserve gametes and embryos in assisted reproductive technology (ART). Our prior research demonstrates that vitrification preserves the viability and functionality of ovarian follicles. However, its impact on follicle-enclosed oocyte remains unknown. The current study investigates whether vitrification, combined with a 3D encapsulated in vitro follicle growth (eIVFG) system, maintains oocyte transcriptome during in vitro follicle development and oocyte maturation. Immature mouse preantral follicles were vitrified and cultured in eIVFG for 8 days to grow to the preovulatory stage, followed with the induction of ovulation and oocyte maturation on day 9, with fresh follicles as the control. Oocytes at germinal vesicle (GV) stage from grown preovulatory follicles on day 8 and oocytes at metaphase II (MII) upon ovulation on day 9 were collected for single-oocyte Smart-Seq2 RNA sequencing analysis. The principal component analysis (PCA) separated GV and MII oocytes into two distinct clusters, but oocytes from fresh and vitrified follicles were largely overlapped. Differential gene expression (DEG) analysis revealed that GV or MII oocytes from fresh and vitrified follicles had comparable expression of maternal effect genes and other genes related to oocyte meiotic and developmental competence. There was a significant transcriptomic change during the GV-to-MII transition. Gene ontology (GO) and KEGG analysis identified DEGs between GV and MII oocytes related to cell cycle, RNA processing, mitochondria, and ribosome. In summary, our study demonstrates that vitrification preserves oocyte transcriptome during in vitro follicle development and oocyte maturation, supporting its potential in fertility preservation. Moreover, our single-oocyte RNA sequencing analysis identifies key DEGs upon GV-to-MII transition, indicating their potential functions in underpinning oocyte meiotic and developmental competence.
2025-04-02 | GSE292062 | GEO
Project description:Despite great improvements in assisted reproductive technology (ARTs), the success of in vitro embryo production remains relatively low. Most of the oocytes used to produce in vitro embryos are recovered from smaller follicles, forming a heterogeneous population that must be matured in vitro. Since the original in vitro maturation (IVM ) experiments (Heilbrunn et al. , 1939) the process by which the most competent oocytes are selected to produce blastocysts remains similar and is still based on morphological aspects of the oocyte cytoplasm and the number of layers and compaction of cumulus cells attached to the oocyte surface (Armstrong, 2001, Coticchio et al. , 2004, Krisher, 2003, Lonergan et al. , 2003). Ovaries from crossbred cows (Bos indicus x Bos taurus) were collected immediately after slaughter and transported to the laboratory in saline solution (0.9% NaCl) supplemented with penicillin G (100 IU/mL) and streptomycin sulfate (100 mg/mL) at 35-37C. The follicles were dissected from the ovarian cortex using scissors, scalpels and tweezers in TCM-199 medium supplemented with Hank’s salts and 10% fetal calf serum (FCS; GIBCO BRL) at 36C. Follicles were measured using a graduated eyepiece (OSM-4; Olympus) and then classified morphologically into two groups according to their diameter: 1.0-3.0 mm or ≥8.0 mm. The criteria used for follicle selection included the presence of extensive and fine vascularization and a shiny and translucent appearance. After follicular rupture, the presence of granulosa cells with a regular and healthy appearance and no free-floating particles in the follicular fluid were also used as selection criteria. Only COCs with a homogeneous granulated cytoplasm and at least three layers of compact of cumulus cells were used in the present study. The selected COCs were transferred to a 50 μL droplet of phosphate-buffered saline (PBS), and the cumulus cells were mechanically removed by repeated pipetting. After cumulus collection, they were transferred to a 0.2-mL tube and centrifuged twice for 2 min at 700 g. The supernatant was removed, and 2μL of RNAlater (Applied Biosystems, Foster City, CA, USA) was added to the pellet, which was then stored at -20C until RNA extraction. For each follicle size group, three replicas of pooled cumulus cells corresponding to 30 oocytes were stored for RNA extraction and subsequent microarray assays, and four independent replicates corresponding to 20 oocytes were stored for RNA extraction for subsequent qPCR assays. During oogenesis, the somatic cells that surround the oocyte proliferate and differentiate into cumulus cells (CCs), which are metabolically coupled to the oocyte, forming the cumulus-oocyte complexes (COCs) (van den Hurk and Zhao, 2005). The CCs remain in strong contact with the oocyte, maintained by cytoplasmic bridges called gap-junctions, as well as through justacrine and paracrine signaling networks (Albertini et al. , 2001, Gilchrist et al. , 2004, Tanghe et al. , 2002, Vozzi et al. , 2001, Webb et al. , 2002). This intense bidirectional communication between the CCs and the oocyte is maintained during all phases of folliculogenesis and is essential for the acquisition of oocyte competence that is required for blastocyst development (Assidi et al., 2008, Fair, 2003, Gilchrist et al., 2004). An informative model to access the level of oocyte competence that has been used by many research groups is follicle size (Donnison and Pfeffer, 2004, Franco et al. , 2013, Lequarre et al. , 2005, Mourot et al., 2006). In a previous study, we showed that oocytes obtained from follicles 1-3 mm in diameter are significantly less competent in producing blastocysts than oocytes obtained from follicles ≥8 mm in diameter (Franco et al., 2013). In the present study, we use the same model to compare the gene expression profile of more than 23,000 bovine transcripts between the CCs obtained from incompetent COCs (1-3 mm) and competent COCs (≥8 mm). We have found substantial differences in the expression of several gene clusters representing distinct metabolic pathways such as energy metabolism, cell signaling, cell cycle, DNA repair, meiosis, and inflammation.
2015-01-27 | E-GEOD-65269 | biostudies-arrayexpress
Project description:Oocyte maturation is a complex process that is regulated by a variety of factors. Mitochondria are a key factor affecting oocyte maturation in vitro. Mitochondrial fission process 1 (MTFP1) is located on the inner mitochondrial membrane and mediates the fission of this membrane. However, the role and mechanism of action of MTFP1 in bovine oocyte maturation are still unclear. We found that MTFP1 is expressed at all stages of bovine oocyte maturation. Moreover, MTFP1 knockdown decreased oocyte maturation efficiency. These observations, combined with our transcriptome sequencing results, showed that MTFP1 knockdown caused mitochondrial dysfunction, impaired nuclear cytoplasmic maturation, promoted mitochondrial fusion, induced mitophagy and decreased oocyte apoptosis. In summary, the inner mitochondrial membrane protein MTFP1 plays a crucial role in bovine oocyte maturation. The results provide a reference and theoretical basis for improving the quality of in vitro oocyte maturation and breeding efficiency in beef cattle
2025-12-09 | GSE311939 | GEO
Project description: Not available
| S-EPMC4809558 | biostudies-literature
Project description:The acquisition of oocyte competence, involving both cytoplasmic and nuclear maturation, is critical for successful fertilization and embryonic development in mammals. Discrepancies between in vivo and in vitro studies underline the potential impact of assisted reproductive technologies (ARTs), such as FSH stimulation and in vitro maturation, on oocyte quality and the molecular integrity of bovine oocytes. These insights underscore the importance of optimizing ART conditions to enhance reproductive success in cattle production. Therefore, this study focuses on the gene expression and DNA methylation dynamics in bovine oocytes from dominant follicles, utilising single-cell bisulfite conversion and RNA sequencing (scG&T), without ovarian stimulation. Samples were collected from synchronized heifers during three time points from 24hr to 2hr before ovulation and submitted to scG&T and compared with a dataset of >120 um in diameter oocytes collected by ovary slicing. Our findings from the single-cell RNA sequencing highlight significant changes in oxidative phosphorylation transcripts, indicating the importance of energy metabolism during oocyte capacitation. Additionally, small changes in global CpG methylation and methylation over genomic features were observed. However, modest modifications in CpG windows suggest a less stable epigenome than previously assumed in fully-grown oocytes. In conclusion, the present study brings new insights into molecular changes in bovine oocytes collected from dominant follicles, which can contribute to oocyte quality improvement in cattle production.
2025-07-14 | GSE297927 | GEO
Project description:oocytes were collected from ovarys of 6 day old female mice. Targets from four biological replicates of each were generated and the expression profiles were determined using Affymetrix MOE430 A & B. Experiment Overall Design: 4 oocyte at primary follicle stage biological replicates were analyzed
2008-06-12 | E-GEOD-3291 | biostudies-arrayexpress
Project description:The palmitoyl-proteome of bovine cumulus-oocyte complexes (COCs) cells was investigated by combining acyl-biotin exchange chemistry and quantitative mass spectrometry analysis.
2022-02-15 | PXD020540 | Pride
Project description:Thecal tissue forms a layer around the follicle just prior to antral stage and grows with the follicle (containing an oocyte) as it matures. The innermost component (theca interna) supplies hormones and other factors necessary to the growth and development of the granulosa and oocyte. Most follicles regress and die (become atretic) at the antral stage, and this process as well as development of the follicle are undoubtedly influenced by the theca. Transcriptional changes in ovarian theca interna at the global level were examined by microarray during atresia and development to improve our understanding of the mechanisms involved.
2014-05-20 | GSE49505 | GEO
Project description:MicroRNAs (miRNA) are important regulators of oocyte maturation, playing a key role in modulating gene expression both in a temporal- and spatial-specific manner. These small non-coding RNAs are involved in important processes during oocyte maturation, acting as messengers between the oocyte and its surrounding cumulus cells. Despite its significance, the bidirectional communication mechanism is still unknown. To test miRNA communication between oocyte and surrounding cumulus cells through the gap junctions the gap junctions were either blocked with carbenoxolone or not. MiRNA sequencing of oocytes at 1, 6, and 22 hours of in vitro maturation was then performed. Among the differentially expressed miRNAs, bta-miR-21-5p, a regulator of cumulus cell viability and oocyte maturation, was the only previously known miRNA. Furthermore, by labeling a bta-miR-21-5p mimic with FAM, crossing of this miRNA through the gap junctions within the cumulus-oocyte complex could be visualized and internalization in the oocyte was confirmed by RT-qPCR. In conclusion, this study provides, for the first time, evidence that miRNA communication within the bovine cumulus-oocyte complex is enabled through the gap junctional network.
2024-08-02 | GSE252748 | GEO