Multi-Omics Analysis of Ovine Atretic Follicles：Linoleic Acid Metabolite 13-Hydroxyoctadecadienoic Acid as a Biphasic Ferroptosis Modulator in Granulosa Cells
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ABSTRACT:
ORGANISM(S): Ovis aries
PROVIDER: GSE292797 | GEO | 2025/03/28
REPOSITORIES: GEO
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Project description:Ovarian follicular atresia is a natural physiological process, but its mechanism is not fully understood. In this study, a quantitative proteomic and phosphoproteomic of granulosa cell (GC) in the healthy (H), slightly atretic (SA), and atretic follicles (A) of porcine were performed by TMT labeling, enrichment of phosphopeptides and LC-MS/MS analysis. Altogether, 6,201 proteins were quantified, and 4,723 phosphorylation sites of 1,760 proteins were quantified. There are 24 (11 up, 13 down) and 50 (29 up, 21 down) proteins with FC>5 in H/SA and H/A, respectively. In addition, there are 20 (H/SA, up) and 39 (H/A, up) phosphosites with FC>7, which could serve as potential biomarkers to distinguish different quality categories of follicles. The results of western blotting and immunofluorescence indicated the reliability of the proteomic analysis. Further analysis of the differential expressed proteins (DEPs) and phosphorylated proteins (DEPPs) revealed some key proteins (e.g. MIF, beta catenin, integrin β2), key phosphosites (e.g. S76 of Caspase6, S22 and S636 of Lamin A/C), pathways (e.g. apoptosis, regulation of actin cytoskeleton pathway), transcription factors (e.g. STAT5A, FOXO1, BCLAF1), and kinases (e.g. PBK, CDK5, CDK12, AKT3) that involved in atresia process. Our proteomic and phosphoproteomic profiling and functional research comprehensively analyze the dynamic changes of protein expression and phosphorylation during follicular atresia, and gave a new explanation for the regulation of this process.
2021-09-09 | PXD020899 | Pride
Project description:Folliculogenesis corresponds to the development of follicles leading to either ovulation or degeneration (a process called atresia). Even if atresia involves an apoptosis process, this mechanism is not well understood. The objective of this experiment was : 1) to analyse gene expression in pig granulosa cells of ovarian follicles during atresia using transcriptome analysis with a 9 024 cDNAs microarray, 2) the identification of gene networks involved in pig ovarian follicular atresia. Granulosa cells were isolated from atretic follicles (small, medium or large). Gene expression was analysed by hybridization of nylon cDNA microarrays. The images were quantified using Bzscan software and the data were managed with BASE software. Statistical analysis was performed using R software. Keywords: pig ovary, folliculogenesis, atresia, gene expression, cDNA microarray, bio-analysis
2008-12-31 | GSE7635 | GEO
Project description:Background: In mammals, female fertility is determined by the outcome of follicular development (ovulation or atresia). Follicular atresia is a complex physiological process that results in the degeneration of oocytes from the ovary. However, the molecular mechanisms of oocyte degeneration and key protein markers of follicular atresia remain unclear. In this study, we explored the complex transcriptional regulatory mechanisms and protein profiles in oocytes and follicular fluid in atretic follicle stages using single-cell RNA sequencing and tandem mass tag proteomics. Results: First, through paired analysis of different follicle development stages, we identified 175 atresia-specific genes and eight candidate oocyte-secreted factors, including PKG1, YTHDF2, and MYC. Meanwhile, we also characterized unique features of the oocyte transcriptional landscape in the atretic follicle stage that displayed cell death-related transcriptional changes and mechanisms, such as autophagy (TBK1 and IRS4), necroptosis (PKR), and apoptosis (MARCKS). Moreover, we identified atresia-specific genes, namely FTH1, TF, and ACSL4, which may participate in regulation of oocyte ferroptosis in atretic follicles through a series of mechanisms including ferritinophagy, ferritin transport, and lipid metabolism. Additionally, we uncovered 333 differentially expressed proteins that may coordinate follicular atresia and revealed key pathways, such as negative regulation of angiogenesis, metabolic pathways, and transcription and mRNA splicing, that lead to oocyte degeneration. Finally, by combining transcriptome and proteomics analyses, we identified two oocyte-secreted biomarkers, PGK1 and ANGPT2, that may be associated with follicular atresia. Conclusions: In conclusion, our work provides a comprehensive characterization of oocyte degeneration in ovine atretic follicles, which provides a basis for establishing an oocyte quality evaluation system and understanding the mechanism of follicular atresia in sheep, as well as an important reference for in vitro production of embryos.
2024-06-05 | GSE268651 | GEO
Project description:Granulosa cells mature and die as ovarian follicles enlarge and die (undergo atresia) under the influence of hormones and intrafollicular factors. Later in follicular development, a fluid-filled antrum is formed, a process which is accompanied by a high rate of atresia. These small antral follicles (5 mm or less in diameter in the cow) contain granulosa of 2 different phenotypes, rounded or columnar, whereas follicles larger than 5 mm have the rounded phenotype only. Prior to ovulation, in larger follicles greater than 10 mm in size, the granulosa begin to migrate and differentiate in preparation for oocyte release and formation of the corpus luteum. These two key phases of follicular development were studied by gene expression microarray analysis using a bovine model to dissect the molecular mechanisms underlying these processes. Four groups of bovine ovarian follicles were selected for analysis. Follicle size, type and array number for each group are as follows: small (3-5 mm) healthy rounded (n=5), small healthy columnar (n=5), atretic (n=5) and large healthy (>10 mm; n=4). For each group, the RNA from a single follicle was used to hybridise an array, except for 3 small healthy samples which were pooled from 2 follicles each due to low RNA.
2014-05-20 | E-GEOD-39589 | biostudies-arrayexpress
Project description:Folliculogenesis corresponds to the development of follicles leading to either ovulation or degeneration (a process called atresia). Even if atresia involves an apoptosis process, this mechanism is not well understood. The objective of this experiment was : 1) to analyse gene expression in pig granulosa cells of ovarian follicles during atresia using transcriptome analysis with a 9 024 cDNAs microarray, 2) the identification of gene networks involved in pig ovarian follicular atresia. Granulosa cells were isolated from atretic follicles (small, medium or large). Gene expression was analysed by hybridization of nylon cDNA microarrays. The images were quantified using Bzscan software and the data were managed with BASE software. Statistical analysis was performed using R software. Keywords: pig ovary, folliculogenesis, atresia, gene expression, cDNA microarray, bio-analysis Calibrated follicles were isolated from porcine ovaries after ovariectomy (24h ou 96h after the end of progestagen treatment: Regumate 18 days, 20 mg/days). The follicular status was determined by histological criteria using Feulgen coloration (Monget et al. 1993, Besnard et al. 1996). Healthy follicle is characterized by frequent mitosis and pyknosis absence in granulosa cells. Presence of frequent pyknotic bodies in granulosa cells and no mitosis were allocated to atretic cells. Two cross-classified biological factors of pig ovarian follicles are considered : 1) the stage of development of the follicular growth : small (1-2 mm of diameter), medium (3 mm) and large (>5 mm) follicles; 2) the follicular status: healthy (S1) vs beginning (A2) or advanced (A3) atresia. Gene expression was analysed by hybridization of nylon cDNA microarrays (GPL3729). After hybridisation the data analysis using BASE software and statistical analysis was performed to identify regulated genes. For each of three follicle class and three follicle status combination 4 biological replicates were considered. Total 36 membranes were first hybridized with a vector oligonucleotide labeled (T7 probe 5'-taatacgactcactataggga-3') with gamma33P ATP at 42°C during 12h to determine the quality of the spotting process. After being washed, the arrays were exposed 6 or 24 hours to radioisotopic-sensitive imaging plates (BAS-2025, Fujifilm, Raytest France S.A.R.L., Courbevoie, France). The imaging plates were scanned thereafter with a phosphor imaging system at a 25 õm resolution (BAS-5000, Fujifilm, Raytest France S.A.R.L., Courbevoie, France). Data were acquired with help of a microarray image processing software - AGScan (Cathelin, 2006, http://mulcyber.toulouse.inra.fr/projects/agscan/). Fixed circle segmentation, i.e. a grid process with a fixed spot diameter was applied. The hybridization images were quantified by the extraction of the intensity for each spot. Arrays images and data were stored using BASE software (Saal LH, Troein C, Vallon-Christersson J, Gruvberger S, Borg A, and Peterson C. BioArray Software Environment (BASE): a platform for comprehensive management and analysis of microarray data. Genome Biol 3: SOFTWARE0003, 2002) adapted by Sigenae (http://www.sigenae.org) for analysis and storage of radioactitive microarray data
2010-06-24 | E-GEOD-7635 | biostudies-arrayexpress
Project description:In mammals, ovarian folliculogenesis leading to the ovulation of completely mature oocytes is a long and complex process that is regulated at different levels. The mechanisms that underlie the selection of one or several dominant follicles as well as the regulation of the number of ovulating follicles are largely unknown. In this project, we proposed to study the genetic determinism that underlies the difference of ovulation rate between species (cattle and pigs), by studying two processes that exist in ovary: follicular development and follicular atresia. Towards this purpose, we made a comparative transcriptomics study on granulosa cells. Pig and cattle comparison was achieved by a transcriptome analysis with a 9K nylon pig microarray (GPL3729) on granulosa cells from either small healthy antral follicles (SHF), small atretic follicles (SAF) or large healthy antral follicles (LHF). The images were quantified using AGscan software and the data were managed with BASE software. Statistical analysis was performed using R software. Transcriptomic analysis on pig and cattle evidenced 997 differentially expressed genes (FDR1%) between the three follicle classes and/or the two species. This research project which implicated three laboratories from INRA: "Laboratoire de Genetique Cellulaire" (UMR444-LGC) , "Station d'Amelioration Genetique des Animaux" (UR 631-SAGA) and "Physiologie des Comportements et de la reproduction" (UMR 85-PRC) benefited from both European funding through SABRE project and French ANR funding through GenOvul project. Keywords: transcriptome analysis, pig, cattle, ovary, folliculogenesis, gene expression, cDNA microarray The data were obtained from 34 RNA samples: 10 small healthy follicles samples (6 for sows and 4 for cows), 13 small atretic follicles samples (7 for sows and 6 for cows) and 11 samples for large healthy follicles (6 for sows and 5 for cows). They were hybridized on a 9K pig nylon microarray (GPL3729).
2012-12-31 | E-GEOD-24221 | biostudies-arrayexpress
Project description:Granulosa cells originating from three different phases of antral follicle growth were compared: growing (G), plateau (P) and atresia (A), as categorized by flow cytometry profiles of DNA. The growing and atretic conditions were each hybridized against the plateau condition as a reference in order to understand the specific biological mechanisms modulated in this class of follicles. Three-condition experiment, Plateau, Growing vs Atretic granulosa cells. Biological replicates: 4 . Dye-swap experiment.
2014-06-02 | E-GEOD-56145 | biostudies-arrayexpress
Project description:Granulosa cells mature and die as ovarian follicles enlarge and die (undergo atresia) under the influence of hormones and intrafollicular factors. Later in follicular development, a fluid-filled antrum is formed, a process which is accompanied by a high rate of atresia. These small antral follicles (5 mm or less in diameter in the cow) contain granulosa of 2 different phenotypes, rounded or columnar, whereas follicles larger than 5 mm have the rounded phenotype only. Prior to ovulation, in larger follicles greater than 10 mm in size, the granulosa begin to migrate and differentiate in preparation for oocyte release and formation of the corpus luteum. These two key phases of follicular development were studied by gene expression microarray analysis using a bovine model to dissect the molecular mechanisms underlying these processes.
2014-05-20 | GSE39589 | GEO
Project description:Goal:Circular RNAs (circRNAs) are thought to play important roles in multiple biological processes including apoptosis. In the ovary granulosa cell apoptosis plays an essential role in the follicular phase of the estrous cycle as it determines if the selected follicles will degenerate or escape this fate and ovulate. Whether circRNAs play a role in granulosa cell apoptosis is at present unknown. By investigating the potential role of circRNAs in the granulosa cell apoptotic cascade we hope to gain novel information regarding the regulation of antral follicular apoptosis using the porcine ovary as a model. Method: Ribosomal-depleted RNA-sequencing was performed to generate circRNA expression profiles from isolated mural porcine granulosa cells of healthy antral (HA) and atretic antral (AA) follicles, respectively. Results:In total, more than 9,632 circRNAs were identified, of which 62 circRNAs were differentially expressed (DE-circRNAs). GO and KEGG analysis of DE-circRNAs showed that apoptosis, cellular responses to stress and cell cycle pathways, were significantly enriched. Next the characteristics of DE-circRNAs, including back-splicing, RNase R resistance and stability were validated, and miRNA binding sites of DE-circRNAs were predicted. Conclusion: Based on these observations, we conclude that aberrantly expressed circRNAs and their targeted genes seem to be associated with granulosa cell apoptosis and thus antral follicular atresia.
2020-08-31 | GSE136589 | GEO
Project description:Granulosa cells originating from three different phases of antral follicle growth were compared: growing (G), plateau (P) and atresia (A), as categorized by flow cytometry profiles of DNA. The growing and atretic conditions were each hybridized against the plateau condition as a reference in order to understand the specific biological mechanisms modulated in this class of follicles.
2014-06-02 | GSE56145 | GEO