Effects of treatment with bovine oviductal extracellular vesicles on the transcriptome of bovine day 7 in vitro produced embryos
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PROVIDER: PRJNA600994 | ENA |
REPOSITORIES: ENA
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Project description:Oviductal extracellular vesicles (oEVs) are emerging as key players in gamete/embryo-oviduct interactions contributing to successful pregnancy. Various positive effects of oEVs on gametes and early embryos have been found in vitro. To find out if these effects are associated with changes of embryonic gene expression, the embryonic transcriptome was analyzed after supplementation of bovine embryos with bovine oEVs during in vitro culture (IVC) in comparison to corresponding controls without oEVs. Embryos were cultured during 7 days of IVC with fresh oEVs, oEVs frozen after isolation from oviductal fluid or without oEVs (control), since a previous study showed different effects of fresh and frozen oEVs. Five pools of 10-14 embryos were analyzed for each group by low-input Illumina RNA-sequencing. Analysis of differential gene expression revealed 221 differentially expressed genes (DEGs) for the comparison of frozen oEVs vs control, 67 DEGs for fresh vs frozen oEVs, and only minor differences for the comparison fresh oEVs vs control (28 DEGs). An integrative analysis with the previously studied mRNA and small ncRNA content of bovine oEVs suggested direct effects of oEVs content on the embryonic transcriptome for the supplementation with frozen oEVs, i.e., increase of transcripts found in higher concentrations in oEVs and decrease of transcripts which are potential targets of miRNAs found in oEVs.
2020-03-21 | GSE143596 | 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:Objective To investigate the influence that oviductal extracellular vesicles from patients with endometriosis exert on early embryo development. Design In vitro experimental study Setting University-affiliated hospital. Patient(s) Women with and without endometriosis who underwent hysterectomy (n = 15 in total). Intervention(s) None. Main Outcome Measure(s) Oviductal extracellular vesicles from patients with endometriosis (oEV-EMT) or without endometriosis (oEV-ctrl) were isolated and co-cultured with two-cell murine embryos for 75 hours. Blastocyst rates were recorded. RNA sequencing was used to identified the differentially expressed genes in blastocysts cultured either with oEV-EMT or with oEV-ctrl. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis were performed to identify potential biological processes in embryos that oEV-EMT affect. The functions of oEV on early embryo development were determined by reactive oxygen species (ROS) levels, mitochondrial membrane potentials, total cell numbers and apoptotic cell proportions. Result(s) Extracellular vesicles were successfully isolated from human Fallopian tubal fluid and their characterizations were described. The blastocyst rates were significantly decreased in the oEV-EMT group. RNA sequencing revealed that oxidative phosphorylation was down-regulated in blastocysts cultured with oEV-EMT. Analysis of oxidative stress and apoptosis at the blastocysts stage showed that embryos cultured with oEV-EMT had increased ROS level, decreased mitochondrial membrane potentials, and increased apoptotic index. Total cell numbers were not influenced. Conclusion(s) Oviductal extracellular vesicles from patients with endometriosis exert a negative influence on early embryo development.
2023-06-02 | GSE225323 | GEO
Project description:The mammalian oviduct is a complex, fibromuscular organ known for its role in orchestrating a series of timely and dynamic changes to suitably support early embryogenesis. Establishing successful reproductive outcomes are largely determined through effective and cohesive communication systems between the transient embryo and the maternal environment. Climate change-induced, Heat stress (HS), is one of the largest single stressors compromising reproductive function. Systemic changes in the redox status of the maternal environment, adversely affect fertilization and early embryonic development, negative causation of HS. Oviductal organoids represent a 3-dimensional, biomimetic model to study the physiological impact of HS on the physiology of the oviduct and the subsequent developing embryo. In this study, we aimed to generate a robust bovine oviductal organoid culture system to decrypt the oviducts' differential response to HS in terms of secreted EV-miRNome, to elucidate the impact of thermal stress on oviductal physiology with subsequent potential impacts on early embryo development.
2023-10-27 | GSE221895 | GEO
Project description:Analysis of the mRNA content of bovine oviductal extracellular vesicles during the estrous cycle
| PRJNA433626 | ENA
Project description:The goal of this experiment was to determine the RNA contents of extracellular vesicles isolated from 3-6mm bovine ovarian follicles.
2017-08-15 | GSE96832 | GEO
Project description:Analysis of the small non-coding RNA content of bovine oviductal extracellular vesicles during the estrous cycle
| PRJNA433714 | ENA
Project description:The mammalian oviduct is a complex, fibromuscular organ known for its role in orchestrating a series of timely and dynamic changes to suitably support early embryogenesis. Establishing successful reproductive outcomes are largely determined through effective and cohesive communication systems between the transient embryo and the maternal environment. Climate change-induced, Heat stress (HS), is one of the largest single stressors compromising reproductive function. Systemic changes in the redox status of the maternal environment, adversely affect fertilization and early embryonic development, negative causation of HS. Oviductal organoids represent a 3-dimensional, biomimetic model to study the physiological impact of HS on the physiology of the oviduct and the subsequent developing embryo. In this study, we aimed to generate a robust bovine oviductal organoid culture system to decrypt the oviducts' differential transcriptomic response to HS, to elucidate the impact of thermal stress on oviductal physiology with subsequent potential impacts on early embryo development.
2023-10-27 | GSE221929 | GEO
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