Dynamics of extracellular vesicle-coupled microRNAs in equine follicular fluid associated with follicle selection
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ABSTRACT:
PROVIDER: PRJNA869321 | ENA |
REPOSITORIES: ENA
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| SRR21038410.fastq.gz | Fastqsanger.gz | |||
| SRR21038411.fastq.gz | Fastqsanger.gz | |||
| SRR21038412.fastq.gz | Fastqsanger.gz | |||
| SRR21038413.fastq.gz | Fastqsanger.gz | |||
| SRR21038414.fastq.gz | Fastqsanger.gz |
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Project description:Seasonal variations of extracellular vesicle-coupled microRNAs in equine follicular fluid
| PRJNA1047927 | ENA
Project description:The mammalian follicular development is a well-orchestrated and complex molecular process characterized by sequences of follicular waves that yield the ovulatory follicle. It has been reported that bi-directional communication between the growing oocyte and the surrounding somatic cells is a hallmark of mammalian follicular development, partially mediated by extracellular vesicles (EVs) present in the follicular fluid (FF). Bioactive molecules such as the miRNAs encapsulated in the EVs mediate this communication within the follicular environment. Here, we aimed to decipher the dynamics of the miRNAs in the EVs of equine FF aspirated in vivo during different stages of follicular development. Hence, all follicles ≥6 mm were ablated on Days 10–11 of the estrous cycle (ovulation = Day 0) and the new induced wave was ultrasound-monitored daily until the largest growing follicle for at least three consecutive days was randomly allocated to one of the five groups (n = 8 follicles/group) namely: predeviation (preDEV; 18–20 mm), deviation (DEV; 22–25 mm), postdeviation (postDEV; 26–29 mm), preovulatory (preOV; 30–35 mm), and impending ovulation (IMP; ~40 mm). The FF was collected from each target group by ultrasound-guided transvaginal follicle aspiration. After removal of cells, cellular debris, and larger particles, EVs were isolated from FF groups (4 biological replicates/group; 0.5 mL FF/replicate) using high-speed ultracentrifugation (120,000xg for 70 min). Isolated EVs were characterized using western blot for EV marker proteins, NanoSight Tracking Analysis, and electron microscopy. MiRNA enriched total RNA was isolated from EVs using an Exosomal RNA Isolation kit and small-RNA library preparation and RNAseq (NextSeq500; Illumina) was performed by Novogene Co, Ltd. Approximately 180 known miRNAs were detected in each group with 148 mutually found in all groups. Among the expressed miRNAs, miR-148a, miR-21, miR-192, and miR-99a were the top highly abundant miRNAs in all groups. Cluster analysis exhibited 15 different expression patterns during follicular development. Among these patterns, a group of 23 miRNAs (including miR-106b, miR-199a-5p, and miR-125a-5p) exhibited a stable expression at the preDev until the postDev stage, a significant increase at the preOV stage, and then a significant decrease at the IMP stage. This miRNA cluster has been found to target genes involved in TGF-signaling and endocytosis pathways. Another interesting pattern with 22 miRNAs (including miR-146b-5p, miR-140, and miR-143) exhibited a sharp reduction in their expression from the preDEV until the preOV stage and was predicted to target genes associated with fatty acid biosynthesis, cell cycle, and oocyte meiosis pathways. In contrast, a group of 16 miRNAs (including miR-483, miR-128, and miR-192) gradually increased in expression from preDEV to the postDEV and then downregulated at the preOV stage. This miRNA cluster targets several signaling pathways, including ErbB, thyroid, and estrogen signaling. In conclusion, this study provides greater insights into the stage-specific expression dynamics of the FF EV-miRNAs during equine follicular development, which may play a role in folliculogenesis in mares.
2023-02-23 | GSE211179 | GEO
Project description:This study was performed to discover the association between postpartum during negative energy balance and the expression of extracellular vesicle-coupled microRNAs signatures in follicular fluid of large follicle in dairy cows. For this, next-generation sequencing to total RNA miRNA was performed.
2019-09-23 | GSE129367 | GEO
Project description:Ovarian follicular fluid (FF) comprises a dynamic milieu whose composition alters under seasonal variation, influencing follicle development and oocyte developmental competence. Various cell subtypes (e.g. granulosa, theca, and cumulus) produce cell-secreted nanoparticles, coined extracellular vesicles (EVs), which contain bioactive regulatory molecules such as microRNAs (miRNAs) known to mediate intrafollicular communication through various forms of signaling and information transfer. Markedly, the hallmark feature of EVs transferring cellular knowledge propagates their appealing nature as direct indicators of cellular status, albeit homeostasis, dysfunction, or disease. Photoperiod alterations influencing the regulatory content of FF-EVs as downstream modifiers remain largely unknown, particularly in seasonal breeding animals. Here, we aimed to unpack the impacts of seasonal variation on the miRNA expression profiles of FF-EVs in the mare. Equine pre-ovulatory follicles were monitored and aspirated using ultrasound-guided transvaginal techniques to obtain follicular fluids during non-breeding (spring anovulatory, SAN) and breeding seasons (spring ovulatory, SOV; summer, SUM; and fall ovulatory, FOV). 97 miRNAs were found to be differentially expressed between groups. Pairwise comparisons revealed specific clusters, including six miRNAs involved in the spring transition (miR-149-200b-206-221-328-615) and many others dysregulated during that of the summer (miR-143-192451- 302b-100, and let 7c), the peak time point for breeding. Bioinformatic analyses led to unveiling significant enrichments in various biological functions, such as transcription factor activity, transcription and transcription regulation, nucleic acid binding, sequence-specific DNA binding, p53 signaling, and post-translational modifications. Ultimately, the miRNA content of seasonally divergent FF-EVs poses as potentially relevant indicators of intrafollicular mechanisms directing seasonal transitions and their functional regulatory roles in the pre-ovulatory stage, governing subsequent ovulation.
2024-10-23 | GSE249220 | GEO
Project description:Innumerable similarities in reproductive cyclicity and hormonal alterations highlight the considerable utility of the mare to study aspects of follicular dynamics and reproductive function in view of the largely constricted, human research subjects. The bi-directional communication between the growing oocyte and the surrounding somatic cells embodies the hallmark of mammalian follicular development, partially mediated by extracellular vesicles (EVs) encapsulated with microRNAs (miRNAs) and present in the follicular fluid (FF). Here, we aimed to decipher the dynamics of the miRNAs in EVs from equine FF aspirated in vivo during different stages of follicular development, namely, predeviation (PreDev; 18-20 mm), deviation (Dev; 22-25 mm), postdeviation (PostDev; 26-29 mm), preovulatory (PreOV; 30-35 mm), and impending ovulation (IMP; ∼40 mm). Approximately 176 known miRNAs were found in all groups with 144 mutually detected among all groups. Cluster analysis exhibited 15 different expression patterns during follicular development. Among these patterns, a group of 22 miRNAs (including miR-146b-5p, miR-140, and miR-143) exhibited a sharp reduction in expression from the PreDev until the PreOV stage. Another cluster of 23 miRNAs (including miR-106b, miR-199a-5p, and miR-125a-5p) exhibited a stable expression pattern at the PreDev stage until the PostDev stage, with a significant increase at the PreOV stage followed by a significant decrease at the IMP stage. In conclusion, this study provides greater insights into the stage-specific expression dynamics of FF EV-miRNAs during equine follicular development, which may propose novel approaches to improve ART and provide new biomarkers to facilitate the assessment of ovarian pathophysiological conditions.
| S-EPMC10321592 | biostudies-literature
Project description:The Objective of this study was to establish global gene expression profiles associated with different stages of dominant follicle development in the horse. This was done by collecting ovaries at different stages of an ovulatory follicular wave, when the dominant follicle reached 22 mm (early dominant, ED), 33 mm (late dominant, LD) and 34 h after an i.v. injection of crude equine gonadotropins (CEG; 15 mg i.v.) when the largest follicle reached > 33 mm (preovulatory stage, PO). RNA was then separately collected from granulosa cells and theca-rich follicular wall fractions and was hybridized to the Agilent Horse Gene Expression Microarray. Gene expression was compared between ED and LD and between LD and PO stages within each of granulosa cells and theca walls. Significantly larger numbers of genes were differentially expressed in granulosa cells than in theca walls throughout development, and between LD and PO follicles than between ED and LF follicles. The most salient features were the downregulation of cell cycle genes in granulosa cells during both the ED-LD and LD-PO transitions and the upregulation of genes associated with inflammation, immunity, extracellular matrix remodelling and protection aganints toxic insult in both GCs and TCs, particularly during the LD-PO transition. Five to nine biological replicates per cell type and developmental stage (total of 34 samples) were used in a single dye experiment. Samples were distributed among slides so that each experimental groupeach was represented at least once in each slide. For each gene, mean normalized intensities (n= 5 biological replicates/group) were compared between follicle stages (ED-LD and LD-PO) within each cell type (GC and TC).
2014-03-15 | E-GEOD-52109 | biostudies-arrayexpress
Project description:Differential expression profiles of extracellular vesicle-coupled miRNA in follicular fluid of cows at different metabolic status
| PRJNA531026 | ENA
Project description:The Objective of this study was to establish global gene expression profiles associated with different stages of dominant follicle development in the horse. This was done by collecting ovaries at different stages of an ovulatory follicular wave, when the dominant follicle reached 22 mm (early dominant, ED), 33 mm (late dominant, LD) and 34 h after an i.v. injection of crude equine gonadotropins (CEG; 15 mg i.v.) when the largest follicle reached > 33 mm (preovulatory stage, PO). RNA was then separately collected from granulosa cells and theca-rich follicular wall fractions and was hybridized to the Agilent Horse Gene Expression Microarray. Gene expression was compared between ED and LD and between LD and PO stages within each of granulosa cells and theca walls. Significantly larger numbers of genes were differentially expressed in granulosa cells than in theca walls throughout development, and between LD and PO follicles than between ED and LF follicles. The most salient features were the downregulation of cell cycle genes in granulosa cells during both the ED-LD and LD-PO transitions and the upregulation of genes associated with inflammation, immunity, extracellular matrix remodelling and protection aganints toxic insult in both GCs and TCs, particularly during the LD-PO transition.
2014-03-15 | GSE52109 | GEO
Project description:Equine fertilization cannot be performed in the laboratory as equine spermatozoa do not cross the oocyte´s zona pellucida in vitro. Hence, a more profound study of equine oviductal fluid (OF) composition at the pre-ovulatory and post-ovulatory stages could help to understand what components are required to achieve fertilization in horses. Our work aimed to elucidate the proteomic composition of equine OF at both stages. To do this, OF was obtained postmortem from oviducts of slaughtered mares ipsilateral to a pre-ovulatory follicle (n = 4) or a recent ovulation (n = 4); the samples were kept at -80 °C until analysis. After protein extraction and isobaric tags for relative and absolute quantification (iTRAQ) labelling, the samples were analyzed by nanoliquid chromatography coupled to tandem mass spectrometry (LC-MS/MS). The analysis of the spectra resulted in the identification of a total of 1173 proteins present in pre-ovulatory and post-ovulatory samples; among these, 691 were unique for Equus caballus. Proteins from post-ovulatory oviductal fluid were compared with proteins from pre-ovulatory oviductal fluid and were categorized as up- (positive log fold change) or down-regulated (negative log fold change); 15 proteins were found to be down-regulated in the post-ovulatory fluid and 154 were up-regulated in the post-ovulatory OF compared to the pre-ovulatory fluid. The identified proteins were related to sperm-oviduct interaction, fertilization, and metabolism, among others. Our data reveal consistent differences in the proteome of equine OF prior and after ovulation, helping to increase our understanding in the factors that promote fertilization and early embryo development in horses.
2022-02-17 | PXD025320 | Pride
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