Estrous cycle impacts microRNA content in extracellular vesicles that modulate bovine cumulus cell transcripts during in vitro maturation
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ABSTRACT:
PROVIDER: PRJNA551973 | ENA |
REPOSITORIES: ENA
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| SRR9622500_1.fastq.gz | Fastqsanger.gz | |||
| SRR9622500_2.fastq.gz | Fastqsanger.gz | |||
| SRR9622501_1.fastq.gz | Fastqsanger.gz | |||
| SRR9622501_2.fastq.gz | Fastqsanger.gz | |||
| SRR9622502_1.fastq.gz | Fastqsanger.gz |
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Project description:Purpose: Extracellular vesicles (EVs) are nanoparticles that can be secreted by different cells, including cells found within the ovarian follicle. Currently, EVs are considered an important form of intercellular communication, since they carry biological contents. The goal of this study was to survey the effects of small EVs from follicles at different estrous cycle stage in bovine cumulus cells. Methods: We used an established model to obtain follicular fluid (FF) at early and late estrous cycle stage according to corpus luteum appearance, corresponding to low and high progesterone (P4) levels, respectively. We collected FF from 3-6 mm follicles and isolated small EVs, which were used as a supplement during in vitro maturation (IVM). Cumulus cells were collected from cumulus-oocyte complexes (COCs) pools and the RNAs were obtained and subjected to RNA sequencing. Results: The results showed that small EVs from different estrous cycle stage are capable to affect transcripts in cumulus cells and modulate different pathways and biological processes related to oocyte maturation, ovulation and immune response. Conclusion: This study demonstrated that small EVs from low and high P4 group impact the RNA profile in cumulus cells after 9 hours of in vitro maturation.
2019-12-02 | GSE133587 | GEO
Project description:The success of the early reproductive events depends on an appropriate communication between gametes/embryos and the oviduct. Extracellular vesicles (EVs) contained in oviductal secretions have been suggested as new players in mediating this crucial cross-talk by transferring their cargo (e.g. proteins, mRNA and small ncRNA) from cell to cell. However, little is known about the oviduct EVs composition and their implications in reproductive success. The objective of our study was to determine the changes of oviductal EVs mRNA content under the hormonal influence during the estrous cycle. Methods: EVs, exosomes and microvesicles, were isolated from bovine oviductal fluid at different stages of the estrous cycle (postovulatory-stage, early luteal phase, late luteal phase and pre-ovulatory stage). Total RNA was isolated and used for the preparation of RNA-seq libraries. RNA-sequencing was performed on an Illumina HiSeq 2500. The obtained sequence reads that passed quality filters were mapped to the bovine genome sequence using Hisat 2. Read counts were calculated with QuasR Qcount and statistical analysis with EdgeR to identify differential mRNA abundance across the different stages of the estrous cycle. Results: RNA-sequencing identified 903 differentially expressed transcripts (FDR<0.001) in bovine oviductal EVs across the estrous cycle. Major differences were found between post-ovulatory and the rest of the stages analyzed.Functional annotation of the differentially abundant mRNAs identified functions related to cilia expression, exosome/vesicles, and many transcripts encoding ribosomal proteins. Conclusions: Our findings represent the first extensive oviductal signature of bovine oviductal EVs mRNA content and contribute to a better understanding of the role of EVs as modulators of gamete/embryo-maternal interactions.
2018-07-31 | GSE110399 | GEO
Project description:The success of the early reproductive events depends on an appropriate communication between gametes/embryos and the oviduct. Extracellular vesicles (EVs) contained in oviductal secretions have been suggested as new players in mediating this crucial cross-talk by transferring their cargo (e.g. proteins, mRNA and small ncRNA) from cell to cell. However, little is known about the oviduct EVs composition and their implications in reproductive success. The objective of our study was to determine the changes of oviductal EVs small ncRNA content under the hormonal influence during the estrous cycle. Methods: EVs, exosomes and microvesicles, were isolated from bovine oviductal fluid at different stages of the estrous cycle (postovulatory-stage, early luteal phase, late luteal phase and pre-ovulatory stage). RNA enriched for short molecules was isolated and used for the preparation of small RNA-seq libraries. RNA-sequencing was performed on an Illumina HiSeq 2500. The obtained sequence reads that passed quality filters were annotated by BLAST alignment to various sets of non-coding and coding RNAs. Read counts were calculated based on the frequency of unique sequences after removal of PCR duplicates. Statistical analysis was done with EdgeR to identify differential small ncRNA abundance across the different stages of the estrous cycle. Results: Small RNA-Seq identified the presence of different types of ncRNAs (miRNAs, rRNA fragments, tRNA fragments, snRNA, snoRNA, and other ncRNAs), which were partially also under hormonal influence. Major differences were found between post-ovulatory and early luteal and the rest of the stages for small ncRNAs. Interesting miRNAs identified in oviductal EVs, miR-34b and miR-449a, showed differential abundance among stages. Dysregulation of these miRNAs has been associated with defective cilia in the oviduct and infertility. Conclusions: Our findings represent the first extensive oviductal signature of bovine oviductal EVs miRNA content and contribute to a better understanding of the role of EVs as modulators of gamete/embryo-maternal interactions.
2018-07-31 | GSE110443 | GEO
Project description:The success of the early reproductive events depends on an appropriate communication between gametes/embryos and the oviduct. Extracellular vesicles (EVs) contained in oviductal secretions have been suggested as new players in mediating this crucial cross-talk by transferring their cargo (e.g. proteins, mRNA and small ncRNA) from cell to cell. Therefore, the objective of the project was to analyze the protein content of EVs (exosomes and microvesicles) contained in the bovine oviductal fluid at different stages of the estrus cycle. For this purpose, pairs of oviducts with their attached ovaries were collected from cyclic cows at a local abattoir under different hormonal regulation along the estrous cycle. Oviducts were classified into one of the 4 stages of the estrous cycle according to the morphology of the corpus lutea(CL) and follicle populations: Stage 1 (S1): showed recently ovulated follicle (d 1 to 4 of estrous cycle); Stage 2 (S2): early luteal development with medium or large or both present (d 5 to 11); Stage 3 (S3): fully functional CL yellow or orange in color (d 11 to 17); or Stage 4 (S4): regressing CL with little vasculature and a large preovulatory follicle present (d 18 to 20). EVs were collected by ultracentrifugation and prepared for mass spectrometry analysis.
2018-07-31 | PXD008851 | Pride
Project description:Mouse oocytes control cumulus cell metabolic processes that are deficient in the oocytes themselves and this delegation is necessary for oocyte development. Oocyte-derived bone morphogenetic factor 15 (BMP15) and growth differentiation factor 9 (GDF9) appear to be key regulators of follicular development. The effect of these factors on cumulus cell function before the preovulatory surge of luteinizing hormone (LH) was assessed by analysis of the transcriptomes of cumulus cells from wildtype (WT), Bmp15-/-, and Bmp15-/- Gdf9+/- double mutant (DM) mice using microarray analysis. The biological themes associated with the most highly-affected transcripts were identified using bioinformatic approaches, IPA and GenMAPP/MAPPFinder. There were 5,332, 7,640, and 2,651 transcripts identified to be significantly changed in the comparisons of Bmp15-/- vs. WT, DM vs. WT, and DM vs. Bmp15-/- respectively by the criteria of FC (fold change) p <0.01. Among theses changed transcripts, 744 were commonly changed in all three pair-wise comparisons, and hence were considered to be the most highly affected transcripts by mutation of Bmp15 and Gdf9. IPA Analyses revealed that metabolism was the major theme associated with the most highly-changed transcripts: glycolysis and sterol biosynthesis were the two most significantly affected pathways. Most of the transcripts encoding enzymes for sterol biosynthesis were down-regulated in both mutant cumulus cells and in WT cumulus cell after oocytectomy. Similarly, there was a reduction of de novo-synthesized cholesterol in these cumulus cells. This suggests that oocytes regulate cumulus cell metabolism, particularly sterol biosynthesis, by promoting the expression of corresponding transcripts. Furthermore, in WT-mice, Mvk, Pmvk, Fdps, Sqle, Cyp51, Sc4mol, and Ebp, which encode enzymes in the sterol biosynthetic pathway, were found to be expressed robustly in cumulus cells, but expression was barely detectable in oocytes. Levels of de novo-synthesized cholesterol were significantly higher in cumulus–enclosed oocytes than denuded oocytes. These results indicate that mouse oocytes are deficient in their ability to synthesize cholesterol and require cumulus cells to provide them with products of the sterol biosynthetic pathway. Oocyte-derived BMP15 and GDF9 may promote this metabolic pathway in cumulus cells as compensation for their own deficiencies. Keywords: Gdf9, Bmp15, cumulus cells, transcriptome, metabolism, sterol biosynthesis, genotype comparison, microarray, ingenuity pathways analyses.
2007-10-22 | GSE7225 | GEO
Project description:The oocyte forms a complex with their somatic cumulus cells within the follicle throughout the preovulatory maturation steps. Cumulus cells support their oocyte not only through mechanical protection but also with a close bidirectional exchange of metabolites. Analysis of the oocytes cumulus gives the opportunity to explore non-invasively oocytal well-being and quality. In vitro maturation (IVM) is the first rate-limiting step in in vitro embryo production. Analysis of protein expression in cumulus cells around this critical step helps to explore the impact of maturation conditions and to examine an influence on maturational competence of the oocyte. The goal of this study was the comparison of the cumulus proteome of oocytes with and without maturational competence matured under in vivo and in vitro conditions. Therefore twenty cumulus samples corresponding to single oocytes were analysed. Half of the samples were matured in vivo and the other half in vitro. For each maturation group, cumulus from oocytes matured successfully (SM; n=5) and failed to mature (FM; n=5) were analysed.
2020-06-30 | PXD016679 | Pride
Project description:Analysis of the mRNA content of bovine oviductal extracellular vesicles during the estrous cycle
| PRJNA433626 | ENA
Project description:The present study was undertaken to discover molecular markers in bovine cumulus cells predictive of oocyte competence and elucidate their functional significance. Differences in RNA transcript abundance in cumulus cells harvested from oocytes of adult versus prepubertal animals (model of poor oocyte quality) were identified by microarray analysis. Four genes of interest encoding for the lysosomal cysteine proteinases cathepsin B, S, K and Z and displaying greater transcript abundance in cumulus cells surrounding oocytes harvested from prepubertal animals were chosen for further investigation. Greater mRNA abundance for such genes in cumulus cells of prepubertal oocytes was confirmed by real time RT-PCR. Elevated transcript abundance for cathepsins B, S and Z was also observed in cumulus cells surrounding adult metaphase II oocytes that developed to the blastocyst stage at a low percentage following parthenogenetic activation, versus those that developed at a high percentage. Functional significance of cumulus cell cathepsin expression to oocyte competence was confirmed by treatment of cumulus oocyte complexes during in vitro oocyte maturation with a cell permeable cysteine proteinase (cathepsin) inhibitor. Inhibitor treatment decreased apoptotic nuclei in the cumulus layer and enhanced development of parthenogenetically activated and in vitro fertilized adult oocytes to the blastocyst stage. Stimulatory effects of inhibitor treatment during meiotic maturation on subsequent embryonic development were not observed when oocytes were matured in the absence of cumulus cells. Results support a functional role for cumulus cell cathepsins in compromised oocyte competence and suggest that cumulus cell cathepsin mRNA abundance may be predictive of oocyte quality. Keywords: Bovine, microarray, cDNA, cumulus cells
2008-03-18 | GSE10819 | GEO
Project description:Progesterone (P4) is predicted to act as a negative regulatory hormone for oocyte maturation events; however, its local effects during follicular development remain poorly understood in bovine. The complex process of oocyte meiosis progression is dependent on cellular communication among follicular cells. Besides, the breakdown of this communication, mainly between cumulus cells and oocyte, through the retraction of cumulus projections connecting these cells can impact oocyte maturation. In our study, we observed that ipsilateral follicles containing high intrafollicular P4 concentrations enhance the expression of proteins in follicular small extracellular vesicles predicted to be involved in the retraction of membrane projections based on actin filaments, such transzonal projections (TZPs). Additionally, we found that contralateral follicles containing low intrafollicular P4 concentrations are predicted to regulate cellular communication pathways inducing the maintenance of membrane projections in cumulus cells. Our functional analysis confirmed that supplementation of cumulus-oocyte complexes during in vitro maturation with P4 at concentration similar to ipsilateral follicles reduces the number of TZPs. In summary, our study underscores a direct association between P4 concentration and cumulus-oocyte interaction, with potential consequences for the acquisition of oocyte competence.
2025-01-14 | GSE260681 | GEO
Project description:Transcriptomes of mouse mural granulosa cells were sequenced to identify transcripts expressed in mural granulosa cells of ovaries. Moreover, transcriptomes of cumulus cells were compared between those of young (2 month-old) and old mice (10 month-old) to assess the effects of ageing on cumulus cells. In addition, transcriptomes of cumulus-oocyte complexes were compared between DBA/2 and (C57BL/6 x DBA/2)F1 mice to assess the strain differences.
2016-04-15 | GSE80326 | GEO