Bos taurus
Ontology highlight
ABSTRACT:
REPOSITORIES: ENA
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| DRR126717.fastq.gz | Fastqsanger.gz | |||
| DRR126718.fastq.gz | Fastqsanger.gz | |||
| DRR126719.fastq.gz | Fastqsanger.gz | |||
| DRR126720.fastq.gz | Fastqsanger.gz | |||
| DRR126721.fastq.gz | Fastqsanger.gz |
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Project description:Oocyte competence is progressively acquired during follicle growth, with large follicles containing more often competent oocytes than small follicles. A competent oocyte is able to mature in vitro, be fertilized, and develop to the blastocyst stage. Extracellular vesicles (EVs) and their cargo micro RNAs (miRNAs) are critical in mediating intercellular communication inside the follicle and facilitating oocyte maturation. Whether follicular EVs are also involved in the acquisition of oocyte competence in the growing follicle remains unclear. Here, we have been using an individual culture system to distinguish follicular fluid and their corresponding bovine oocytes as competent or noncompetent. We used a qEV single-column method to isolate EVs from follicular fluid. RNA sequencing revealed 16 differentially expressed (DE) miRNAs in EVs from follicular fluid derived from either competent or noncompetent oocytes . Among the up-regulated miRNAs, we selected bta-miR-34c to further validate its effect on oocyte competence during in vitro maturation using mimics and inhibitors. By supplementing miR-34c mimics to the oocyte maturation medium, we could significantly improve blastocyst quality, as evidenced by higher cell numbers, while supplementation of miR-34c inhibitors produced the opposite effect. Taken together, the up-regulation of miR-34c in EVs derived from follicular fluid from competent oocytes is indicative of its regulatory role, and, when added during in vitro maturation of unselected oocytes, is able to increase total cell number and inner cell mass of resulting embryos, thus contributing to the development of healthy offspring
2024-09-03 | GSE250281 | GEO
Project description:Germ cells of most animals critically depend on piRNAs and Piwi proteins. Surprisingly, piRNAs in mouse oocytes are relatively rare and dispensable. We present compelling evidence for strong Piwi-piRNA expression in oocytes of other mammals. Human fetal oocytes express PIWIL2 and transposon-enriched piRNAs. Oocytes in adult human ovary express PIWIL1 and PIWIL2, while those in bovine ovary just express PIWIL1. In human, macaque and bovine ovaries we find piRNAs that resemble testis-borne pachytene piRNAs. Isolated bovine follicular oocytes were shown to contain abundant, relatively short piRNAs that preferentially target transposable elements. Using label-free quantitative proteome analysis we show that these maturing oocytes strongly and specifically express the thus-far uncharacterized PIWIL3 protein, alongside other known piRNA-pathway components. In bovine early embryos these piRNAs are still abundant, revealing a potential impact of piRNAs on mammalian embryogenesis. Our results reveal unexpected, highly dynamic piRNA pathways in mammalian oocytes and early embryos. Analyses of multiple small RNA libraries obtained from fetal/adult oocytes, cumulus cells, ovary, testis and 2-4 cell stage ivf embryos of multiple mammalian species.
2015-03-27 | E-GEOD-64942 | biostudies-arrayexpress
Project description:Exosomes and microvesicles (i.e., extracellular vesicles; EVs) have been identified within ovarian follicular fluid, and recent evidence suggests that EVs are able to elicit profound effects on ovarian cell function. While existence of miRNA within EVs has been reported, it remains unknown if EV size and concentration as well as their cargos (i.e., proteins and RNA) change during antral follicle growth. Extracellular vesicles isolated from follicular fluid of small, medium and large bovine follicles were similar in size, while concentration of EVs decreased progressively as follicle size increased. Electron microscopy indicated a highly purified population of the lipid bilayer enclosed vesicles that were enriched in exosome biomarkers including CD81 and Alix. Small RNA sequencing identified a large number of known and novel miRNAs that changed in the EVs of different size follicles. Ingenuity Pathway Analysis (IPA) indicated that miRNA abundant in small follicle EV preparations were associated with cell proliferation pathways, while those miRNA abundant in large follicle preparations were related to inflammatory response pathways. These studies are the first to demonstrate that EVs change in their levels and makeup during antral follicle development and point to the potential for a unique vesicle-mediated cell-to-cell communication network within the ovarian follicle. Examination of small RNA population in bovine follicular fluid extracellular vesicles isolated from antral follicles
2016-02-09 | E-GEOD-74879 | biostudies-arrayexpress
Project description:Oocyte competence for early embryo development relies on intercellular communication between the maturing oocyte and preovulatory follicle. Preovulatory follicle maturity, as indicated by serum estradiol concentration or follicle diameter, has previously been linked to pregnancy, follicular fluid metabolites, cumulus-oocyte metabolism, and oocyte competency for embryo development. Such relationships indicate metabolic and developmental programming of the oocyte based on the preovulatory follicle’s physiological status, but downstream impacts on the molecular signature of blastocysts have not been examined. We hypothesized that supplementing maturing oocytes with follicular fluid originating from preovulatory follicles of greater or lesser maturity would impact the transcriptome of resulting blastocysts and indicate metabolic programming of the embryo that originated from the oocyte’s maturation environment. The objective was to investigate the effect of follicle maturity on metabolic preparation of the oocyte by examining the functional genome of blastocysts originating from oocytes matured in the presence of follicular fluid from preovulatory follicles of greater or lesser maturity. In vitro maturing oocytes were supplemented with follicular fluid collected from preovulatory follicles of greater or lesser maturity. Following identical embryo culture procedures, RNA-sequencing was performed on pools of 2 blastocysts (Greater, n = 12; Lesser, n = 15; all with stage code = 7 and quality code = 1). A total of 12,310 gene transcripts were identified in blastocysts after filtering to remove lowly abundant transcripts. There were 113 transcripts that differed in abundance between blastocysts originating from oocytes matured in greater versus lesser maturity follicular fluid (eFDR < 0.01). Although no pathways were significantly enriched with differentially abundant transcripts, transcriptome profiles suggested improved Wnt/β-catenin signaling, metabolism, and protection from oxidative stress in blastocysts derived from oocytes matured in greater maturity follicular fluid, while unregulated cell growth presented in blastocysts resulting from the lesser follicle maturity treatment.
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Project description:Mono(2-ethylhexyl) phthalate (MEHP), the main di(2-ethylhexyl) phthalate (DEHP) metabolite, is a known reproductive toxicant. Residual levels of 20 nM MEHP have been found in follicular fluid aspirated from IVF-treated women and DEHP-treated animals. It is not yet clear whether these residual MEHP levels have any effect on the follicle-enclosed oocyte or developing embryo. To clarify this point, bovine oocytes were matured with or without 20 nM MEHP for 22 h. Microarray analysis was performed for both mature oocytes and 7-day blastocysts. A feasibility examination was performed on mature oocytes (n = 200/group) to reveal a possible direct effect on the oocyte proteomic profile. Transcriptome analysis revealed MEHP-induced alterations in the expression of 456 and 290 genes in oocytes and blastocysts, respectively. The differentially expressed genes are known to be involved in various biological pathways, such as transcription process, cytoskeleton regulation and metabolic pathway. Among these, the expression of 9 genes was impaired in both oocytes exposed to MEHP (i.e., direct effect) and blastocysts developed from those oocytes (i.e., carryover effect). In addition, 191 proteins were found to be affected by MEHP in mature oocytes. The study explores, for the first time, the risk associated with exposing oocytes to physiologically relevant MEHP concentrations to the maternal transcripts. Although it was the oocytes that were exposed to MEHP, alterations carried over to the blastocyst stage, following embryonic genome activation, implying that these embryos are of low quality.
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Project description:Germ cells of most animals critically depend on piRNAs and Piwi proteins. Surprisingly, piRNAs in mouse oocytes are relatively rare and dispensable. We present compelling evidence for strong Piwi-piRNA expression in oocytes of other mammals. Human fetal oocytes express PIWIL2 and transposon-enriched piRNAs. Oocytes in adult human ovary express PIWIL1 and PIWIL2, while those in bovine ovary just express PIWIL1. In human, macaque and bovine ovaries we find piRNAs that resemble testis-borne pachytene piRNAs. Isolated bovine follicular oocytes were shown to contain abundant, relatively short piRNAs that preferentially target transposable elements. Using label-free quantitative proteome analysis we show that these maturing oocytes strongly and specifically express the thus-far uncharacterized PIWIL3 protein, alongside other known piRNA-pathway components. In bovine early embryos these piRNAs are still abundant, revealing a potential impact of piRNAs on mammalian embryogenesis. Our results reveal unexpected, highly dynamic piRNA pathways in mammalian oocytes and early embryos.
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Project description:Bovine follicular fluid extracellular vesicles RNA contents
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Project description:The quality of bovine oocytes is crucial for in vitro embryo production technology, thus it is essential to study the mechanisms of oocyte maturation. N6-methyladenosine (m6A) modification plays a key role in gametogenesis and embryonic development. However, the dynamic landscape and functional roles of m6A modification during bovine oocyte maturation remain unclear. We conducted transcriptome-wide m6A landscape analysis on bovine oocytes at the GV and MII phases using MeRIP-seq and RNA-seq technologies. We identified a total of 6,323 differential m6A peaks in the two groups of oocytes, with 4,422 m6A peaks being significantly upregulated and 1,901 m6A peaks significantly downregulated. The combined analysis of MeRIP-seq and RNA-seq identified 262 genes with significant differences in both methylation modification and transcriptome levels. GO and KEGG enrichment analyses of these genes revealed significant enrichment in biological processes and signaling pathways such as endocytosis, thermogenesis, ribosome, cellular metabolism, Apelin signaling pathway, and HIF-1 signaling pathway. Finally, we examined the roles of m6A modification in bovine oocyte maturation. The results showed that inhibition of m6A modification affects nuclear and cytoplasmic maturation of oocytes. Our results reveal the m6A modification profile during bovine oocyte maturation. Furthermore, we found that inhibition of m6A modification leads to meiotic failure. These results provide a theoretical basis for studying the function of RNA modification in oocyte maturation.
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