Gene expression in bovine ovarian follicle granulosa
Ontology highlight
ABSTRACT:
ORGANISM(S): Bos taurus
PROVIDER: GSE39589 | GEO | 2014/05/20
SECONDARY ACCESSION(S): PRJNA171166
REPOSITORIES: GEO
Similar Datasets
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:Thecal tissue forms a layer around the follicle just prior to antral stage and grows with the follicle (containing an oocyte) as it matures. The innermost component (theca interna) supplies hormones and other factors necessary to the growth and development of the granulosa and oocyte. Most follicles regress and die (become atretic) at the antral stage, and this process as well as development of the follicle are undoubtedly influenced by the theca. Transcriptional changes in ovarian theca interna at the global level were examined by microarray during atresia and development to improve our understanding of the mechanisms involved. Four groups of bovine ovarian follicles were selected for analysis . Follicle size, type and array number for each group are, small (3-5 mm) healthy rounded (n=5), healthy columnar (n=5), atretic(n=5) and large healthy (>9mm), (n=4). For each group, the RNA from the theca interna of a single follicle was used to hybridise an array.
2014-05-20 | E-GEOD-49505 | 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: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:Antral follicle size might be a valuable additive predictive marker for IVF outcome. However, while some studies show positive relations between follicle size and reproductive outcome, others have not been successful in establishing this relation. To better understand consequences of antral follicle size for reproductive outcome, we aimed to obtain insight in follicle size-related granulosa cell processes, as granulosa cells play an essential role in follicular development via the production of growth factors, steroids and metabolic intermediates, needed for follicular growth and oocyte development. Using pigs as a model, we compared gene and protein expression in granulosa cells of smaller and larger follicles in the healthy antral follicle pool at the start of the follicular phase of the estrous cycle. In sows, the early antral follicle pool is very heterogeneous when e.g. size and steroid content of the follicular fluid are considered. To which extent this variety contributes to the developmental competence of the follicles is not clear. Therefore, sows with high variation in antral follicle size (HighVAR) as well as sows with low variation in antral follicle size (LowVAR) were used. Granulosa cells of smaller antral follicles in the healthy antral follicle pool show increased cell proliferation, which was accompanied by a metabolic shift towards aerobic glycolysis (i.e. the Warburg effect), similar to other highly proliferating cells. High granulosa cell proliferation rates in smaller follicles might be regulated via increased granulosa cell expression of AR and EGFR which are activated in response to locally produced mitogens. While granulosa cells of smaller follicles in the pool were more proliferative, indicative of higher follicular growth, granulosa cells of larger follicles in the pool showed less proliferation and were more differentiated, as they showed a higher expression of follicular maturation marker IGF1 and ANGPT1. Our results imply that the inclusion of strict criteria of antral follicle size in IVF protocols might improve reproductive outcome. In addition, we have granulosa cell gene expression of healthy follicles to unravel underlying mechanisms of differences in COC morphology. We compared gene expression in sows with low vs. high-COC-health and found a decreased expression of genes involved in ovarian steroidogenesis (e.g. CYP19A1, ADM, SPP1) and higher expression of genes involved in follicular atresia (e.g. GADD45A, INHBB) in sows with low-COC-health. Thereby we have identified several genes which may serve as markers for follicle developmental competence.
2019-08-15 | GSE125189 | GEO
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: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:Prenatal testosterone treated sheep, similar to PCOS women, manifest reduced cyclicity, functional hyperandrogenism and polycystic ovary (PCO) morphology. The PCO morphology results from increased follicular recruitment and persistence of antral follicles, consequence of reduced follicular growth and atresia, and driven by cell-specific gene expression changes that are poorly understood. Therefore, utilizing RNA sequencing, cell-specific transcriptional changes were assessed in laser capture microdissection isolated antral follicular granulosa and theca cells from 21 months-of- age control and prenatal testosterone treated sheep.
2020-06-18 | GSE150236 | GEO
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: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