ERK1/2-dependent gene expression in the bovine ovulating follicle
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ABSTRACT:
PROVIDER: PRJNA497853 | ENA |
REPOSITORIES: ENA
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| SRR8085101_1.fastq.gz | Fastqsanger.gz | |||
| SRR8085101_2.fastq.gz | Fastqsanger.gz | |||
| SRR8085102_1.fastq.gz | Fastqsanger.gz | |||
| SRR8085102_2.fastq.gz | Fastqsanger.gz | |||
| SRR8085103_1.fastq.gz | Fastqsanger.gz |
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Project description:Purpose: establishing a role for ERK1/2 in bovine ovulation by using RNA-Seq to compare granulosa cells collected from the dominant follicle at different times post GnRH treated with PD0325901 (ERK1/2 signaling inhibitor) Methods: The dominant follicle of cattle was subject to ultrasound-guided intrafollicular injection of either a Vehicle treatment or PD0325901 inhibitor treatment. Thirty minutes later we collected granulosa cells from Vehicle treated cows (Vehicle_0h_GnRH), while other cows were administered with an intramuscular injection of goonadotropin releasing hormone (GnRH) to simulate an LH surge. Six hours later granulosa cells were collected from Vehicle treated and PD0325901 treated cows (Vehicle_6h_GnRH & PD0325901_6h_GnRH). All granulosa cell RNA was subject to the Illumina HiSeqq 2500 PE 125. Quality check for reads was performed using FastQC v0.11.5 and adapters were removed by Trimmomatic v0.36. The data was aligned to the bovine Genome (Btau4.6.1) using Bowtie v2.2.1.0 and TopHatv2.0.11. Next, the rnumber of reads mapped to each gene were counted using HTSeq v0.6.1 Results: The counts from our 8 samples (Vehicle_0h_GnRH; n=2 , Vehicle_6h_GnRH; n=3, PD0325901_6h_GnRH; n=3) were subject to analysis to determine differentially expressed genes (DEGs) between our groups. Using The DESeq2 R package we observed 2 121 DEGs between Vehicle_0h_GnRH & Vehicle_6h_GnRH and 285 DEGs between Vehicle_6h_GnRH and PD0325901_6h_GnRH. We confirmed our RNA-seq analysis for a number of known genes in ovulation (ADAMTS1, TNFAIPs, EGR1, etc..) by qPCR. Conclusions: ERK1/2 is essential for ovulation in cattle as observed by the DEGs in granulosa cells in the pre-ovulatory follicle. To our knoweldge, we are the firs tto iinhbiti ERK1/2 signaling in vivo in cattle.
2018-10-23 | GSE121588 | GEO
Project description:Ovulation is triggered by gonadotropin surge-induced signaling cascades. To study the role of extracellular signal-regulated kinase 1/2 (ERK1/2) in bovine ovulation, we administered the pharmacological inhibitor, PD0325901, into the preovulatory dominant follicle by intrafollicular injection. Four of five cows treated with 50?µM PD0325901 failed to ovulate. To uncover the molecular basis of anovulation in ERK1/2-inhibited cows, we collected granulosa and theca cells from Vehicle and PD0325901 treated follicles. Next-generation sequencing of granulosa cell RNA revealed 285 differentially expressed genes between Vehicle and PD0325901-treated granulosa cells at 6?h post-GnRH. Multiple inflammation-related pathways were enriched among the differentially expressed genes. The ERK1/2 dependent LH-induced genes in granulosa cells included EGR1, ADAMTS1, STAT3 and TNFAIP6. Surprisingly, PD0325901 treatment did not affect STAR expression in granulosa cells at 6?h post-GnRH. Granulosa cells had higher STAR protein and theca cells had higher levels of STAR mRNA in ERK1/2-inhibited follicles. Further, both granulosa and theca cells of ERK1/2-inhibited follicles had higher expression of SLC16A1, a monocarboxylate transporter, transporting substances including ?-hydroxybutyrate across the plasma membrane. Taken together, ERK1/2 plays a significant role in mediating LH surge-induced gene expression in granulosa and theca cells of the ovulating follicle in cattle.
| S-EPMC6212447 | biostudies-literature
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: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:The objective of the study was to compare the transcriptome of the bovine oocyte coming from distinct follicle sizes.
2015-11-15 | GSE48283 | GEO
Project description:A key transition in ovarian follicular development is the activation of resting primordial follicles to the growing primary follicle stage. The signals that regulate activation are still not well understood, especially in non-rodent species. we combined a microarray approach with an in vitro system to gain insight into the regulation of follicle activation. Genes and pathways identified in this study provide interesting candidates for further investigation of mechanisms underlying follicle activation. Fetal bovine ovarian cortical pieces enriched for primordial or primary follicles were obtained by culture with cotrol medium supplemented with TS+ (transferrin, selenous acid, BSA, and linoleic acid) or with insulin (ITS+), a factor that stimulate bovine follicle activation. Total RNA was extracted. Differences in global gene expression profiles between pieces enriched for primordial or primary follicles were determined by microcarray analysis.
2015-10-31 | E-GEOD-69094 | 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.
2014-05-20 | GSE49505 | 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.
2014-05-20 | GSE39589 | GEO
Project description:The objective was to determine age-associated changes in the transcriptome of granulosa cells recovered from the dominant follicle at the time of selection. Granulosa cells were collected from the dominant follicle of aged and young cows after ovariectomy (15 ±1.5 years, n=3 and 6 ± 1.1 years, n=3), or ultrasound-guided follicle aspiration (16±2.1 years, n=4 and 7±1.2 years, 6 ± 1.1 years, n=4) 3-days after ovulation (Day 0). Messenger-RNA was extracted, amplified, labeled with florescent dyes, and hybridized with bovine-specific microarrays (GEO accession # GPL13226). Target intensities were analyzed to determine differential gene expression in granulosa cells from aged vs. young cows. A total 169 genes were differentially expressed (≥ 2 fold-change; P≤0.05) between age groups. In conclusion, granulosa cells collected at the time of selection of the dominant follicle exhibited age-related changes in the transcriptome that may explain follicle-associated loss of oocyte competence in aged cows.
2016-03-31 | GSE52480 | GEO
Project description:The objective was to determine age-associated changes in the transcriptome of granulosa cells recovered from the dominant follicle at the time of selection. Granulosa cells were collected from the dominant follicle of aged and young cows after ovariectomy (15 ±1.5 years, n=3 and 6 ± 1.1 years, n=3), or ultrasound-guided follicle aspiration (16±2.1 years, n=4 and 7±1.2 years, 6 ± 1.1 years, n=4) 3-days after ovulation (Day 0). Messenger-RNA was extracted, amplified, labeled with florescent dyes, and hybridized with bovine-specific microarrays (GEO accession # GPL13226). Target intensities were analyzed to determine differential gene expression in granulosa cells from aged vs. young cows. A total 169 genes were differentially expressed (⥠2 fold-change; Pâ¤0.05) between age groups. In conclusion, granulosa cells collected at the time of selection of the dominant follicle exhibited age-related changes in the transcriptome that may explain follicle-associated loss of oocyte competence in aged cows. Granulosa cells of the dominant follicle at the time of selection (aged vs.young cows). Three biological replicates (each composed of one aged and one young cow). 3 Three technical replicate (dye swap). One biological or technical replicate per array.
2016-03-31 | E-GEOD-52480 | biostudies-arrayexpress