Cellular and extracellular miRNA profiles of pre-ovulatory follicles indicate signalling disturbances in polycystic ovaries
Ontology highlight
ABSTRACT:
PROVIDER: PRJNA659926 | ENA |
REPOSITORIES: ENA
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| SRR12539417.fastq.gz | Fastqsanger.gz | |||
| SRR12539418.fastq.gz | Fastqsanger.gz | |||
| SRR12539419.fastq.gz | Fastqsanger.gz | |||
| SRR12539420.fastq.gz | Fastqsanger.gz | |||
| SRR12539421.fastq.gz | Fastqsanger.gz |
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Project description:Cell-free RNAs have been used as diagnostic disease markers describing the state of tissue environment. The origin and potential functions of such RNAs in human pre-ovulatory follicle, the environment of oocyte maturation, are unclear. We aimed to describe the small RNA profiles of three follicular compartments of the same follicle: mural granulosa cells (MGC), cell-free follicular fluid (FF) and extracellular vesicles (EV) of the FF. Focusing on microRNAs, their signatures and regulated pathways were compared between infertile women with polycystic ovaries and fertile control group.
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Project description:Cell-free RNAs have the potential to act as a means of gene expression regulation between cells and are therefore used as diagnostic markers describing the state of tissue environment. The origin and functions of such RNAs in human ovarian follicle, the environment of oocyte maturation, are unclear. The current study investigates the difference in the microRNA profiles of fertile women and polycystic ovary syndrome (PCOS) patients in three compartments from the same preovulatory follicle: mural granulosa cells (MGC), cell-free follicular fluid (FF), and extracellular vesicles (EV) of the FF by small RNA sequencing. In silico analysis was used for the prediction and over-representation of targeted pathways for the detected microRNAs. PCOS follicles were distinguished from normal tissue by the differential expression of 30 microRNAs in MGC and 10 microRNAs in FF (FDR < 0.1) that commonly regulate cytokine signaling pathways. The concentration of EV-s was higher in the FF of PCOS patients (p = 0.04) containing eight differentially expressed microRNAs (p < 0.05). In addition, we present the microRNA profiles of MGC, FF, and EV in the fertile follicle and demonstrate that microRNAs loaded into EVs target mRNAs of distinct signaling pathways in comparison to microRNAs in FF. To conclude, the three follicular compartments play distinct roles in the signaling disturbances associated with PCOS.
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Project description:To investigate the difference between the pre-ovulatory F5 and pre-recruitment follicular stages in laying hens and identify master regulators that control the processes in these two follicular stages. RNA was isolated from a group of 3-5 pre-recruitment follicles and the most recently recruited pre-ovulatory follicle (F5). After RNA isolation, follicles underwent RNA sequencing and differential gene expression analysis occurred.
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Project description:We deep sequenced and analyzed circRNA using deep RNA sequencing (RNA-seq) in pre-ovulatory follicle samples of Macheng black goats and Boer goats. We analyzed the RNA-seq data with 301 million reads and 288 million reads, and reveal the expression profiles of circRNAs and predicted 13,950 circRNAs. 827 circRNA host genes, mostly related to transferase activity and metabolic process. Twenty-four circRNAs were upregulated and 13 were downregulated in the pre-ovulatory follicles of the Boer group compared to their expression in the Macheng group.
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Project description:Experiments were designed to evaluate changes in the transcriptome (mRNA levels) in the ovulatory, luteinizing follicle of rhesus monkeys, using a controlled ovulation (COv) model that permits analysis of the naturally selected, dominant follicle at specific intervals (0, 12, 24, 36 hours) after exposure to an ovulatory (exogenous hCG) stimulus during the menstrual cycle. Total RNA was prepared from individual follicles (n=4-8/timepoint), with an aliquot used for microarray analysis (AffymetrixTM Rhesus Macaque Genome Array) and the remainder applied to quantitative real-time PCR (q-PCR) assays. The microarray data from individual samples distinctly clustered according to timepoints, and ovulated follicles displayed markedly different expression patterns from unruptured follicles at 36 h. Between timepoint comparisons revealed profound changes in mRNA expression profiles. The dynamic pattern of mRNA expression for steroidogenic enzymes (CYP17A, CYP19A, HSD3B2, HSD11B1, HSD11B2), StAR, and gonadotropin receptors (LHCGR, FSHR) as determined by microarray analysis correlated precisely with those from blinded q-PCR assays. Patterns of mRNA expression for EGF-like factors (AREG, EREG) and processes (HAS2, TNFAIP6) implicated in cumulus-oocyte maturation/expansion were also comparable between assays. Thus, several mRNAs displayed the expected expression pattern for purported theca (e.g., CYP17A, AREG), granulosa (CYP19A, FSHR), cumulus (HAS2, TNFAIP6) cell, and surface epithelium (HSD11B) related genes in the rodent/primate preovulatory follicle. This database will be of great value in analyzing molecular and cellular pathways associated with periovulatory events in the primate follicle (e.g. follicle rupture, luteinization, inflammatory response, and angiogenesis), and for identifying novel gene products controlling mammalian fertility. Keywords: time course Total RNA was prepared from individual follicles (n=4-8/timepoint). Dominant follicles were selected specific intervals (0, 12, 24, 36 hours) after exposure to an ovulatory (exogenous hCG) stimulus during the menstrual cycle.
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Project description:Human ovarian follicles develop a fluid-filled antrum when they reach a diameter of around 250 µm. The antrum contains follicular fluid (FF) composed of secretions from cells in the follicle and from circulation. The composition of FF undergoes massive changes in relation to follicular development. Previously, only FF from large pre-ovulatory follicles aspirated just prior to ovulation, has been analyzed by proteomics; resulting in a high proportion of plasma proteins being detected due to diffusion of proteins through the increasingly leaky basal membrane in larger follicles. We present the first proteomics analysis of FF from normal unstimulated human small antral ovarian follicles (diameter: 3-8mm) identifying the largest number of proteins reported in human FF to date: 2,050 proteins of which 1,151 were identified for the first time in human FF by mass spectrometry. Furthermore, an identified growth factor, midkine, was shown to impact follicular regulatory processes including enhancement of nuclear maturation of immature human oocytes.
2021-09-08 | PXD013344 | Pride
Project description:Protein composition of human ovarian follicular fluid (FF) constitutes the microenvironment for oocyte development. Several proteomics studies of FF from pre-ovulatory follicles have revealed insights on oocyte maturation, however, there is a lack of knowledge on changes produced at protein levels in the FF of human small antral follicles (hSAF) related to the upcoming oocyte maturation. Using mass spectrometry-based proteomics, we evaluated the protein composition of FF that surrounds oocytes capable to reach metaphase II (MII) after IVM with the protein profile of FF that surrounds immature oocytes. The samples were collected from small antral follicles (size 6.0 ± 1.5 mm) extracted from six women, from which two or three samples were extracted. The comparison was based on both, a multivariate (sPLS-DA) and univariate analyses (t-test).
2021-09-09 | PXD021338 | Pride
Project description:Leukocytes, known as key effectors for ovulation, were found to specifically express oncostatin-M (OSM), while its receptors (OSMR and IL6ST) were abundant in granulosa cells, as shown in single-cell transcriptomics of follicular aspirates. OSM levels in the follicular fluid ranged 20.99 to 429.81 pg/ml. Increases in OSM-positive cells and OSMR and IL6ST expression were detected in ovulatory follicles post-hCG administration. Treating recombinant human OSM protein (rhOSM) to hCG-primed human granulosa/lutein cells activated signaling mediators, e.g., STATs. This treatment enabled hGLC cultures to mimic the in vivo ovulatory steroidogenesis dynamic, characterized by increased progesterone and reduced estradiol, unlike hCG alone. Additionally, rhOSM modulated gene expression associated with various ovulatory changes. This evidence suggests that leukocyte-derived OSM is a groundbreaking factor in human ovulation.
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Project description:The molecular mechanisms that regulate the pivotal transformation processes observed in the follicular wall following the pre-ovulatory LH-surge, are still not established, particularly for cells of the thecal layer. To elucidate thecal and granulosa cell type-specific biological functions and signaling pathways, large dominant bovine follicles were collected before and 21 hrs after an exogenous GnRH induced LH surge. Because LH receptor density varies within the granulosa cell populations, antral granulosa (aGC; those aspirated by follicular puncture) and membrane associated granulosa (mGC; those scraped from the follicular wall) were compared to thecal cell expression profiles determined by mRNA microarrays. Thecal cell gene expression was less affected in the peri-ovulatory follicle when compared to granulosa cells, as evidenced by only 2% versus 25% of the ~11,000 genes expressed changing in response to the LH surge, respectively. The majority of the 203 LH-regulated thecal genes were also LH regulated in granulosa cells, leaving a total of 58 genes as LH-regulated theca cell specific genes. Most of the 58 genes (i.e., 74%) thecal specific genes including several known thecal markers (CYP17A1, NR5A1) were downregulated, while most genes identified are new to theca. Many of the newly identified upregulated thecal genes (e.g., PTX3, RND3, PPP4R4) were also upregulated in granulosa. Minimal expression differences were observed between aGC and mGC, however, transcripts encoding extracellular proteins (NID2) and matrix modulators (ADAMTS1, SASH1) predominated these differences. We also identified large numbers of unknown LH-regulated granulosa cell genes and discuss their putative roles in ovarian function.
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