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Gene expression analysis of bovine oocytes with high developmental competence obtained from FSH-stimulated animals

ABSTRACT: The objective of the study was to analyze the impact of various lenght FSH withdrawal period (coasting) after the last FSH injection on transcriptome changes in in vivo bovine oocytes at the GV stage. Oocytes were collected from super-stimulated animals after 20, 44, 68 and 92 hours of coasting. For each cow individually (biological replicate), each coasting time was compared to others (i.e. 20 hrs vs 44 hrs; 20 hrs vs 68 hrs; 20 hrs vs 92 hrs; 44 hrs vs 68 hrs; 44 hrs vs 92 hrs and 68 hrs vs 92 hrs) for a total of six comparisons. Overall, 36 hybridizations, corresponding to the three cows and six comparisons were done using a dye-swap set-up.

ORGANISM(S): Bos taurus

SUBMITTER: Remi Labrecque

PROVIDER: E-GEOD-38345 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Gene expression analysis of bovine oocytes with high developmental competence obtained from FSH-stimulated animals.

Labrecque Rémi R Vigneault Christian C Blondin Patrick P Sirard Marc-André MA

Molecular reproduction and development 20130424 6

Recent progress in the ovarian stimulation protocol used for bovine in vitro maturation and fertilization, especially through optimization of the follicle-stimulating hormone (FSH) withdrawal period ("coasting") after ovarian pre-treatment with FSH, has significantly improved blastocyst outcome. Despite this important success, the underlying factors leading to improved oocyte quality have not yet been identified. The aim of this project was to compare the transcriptome of germinal vesicle-stage ...[more]

PMID: 23559376

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Cumulus cell gene expression associated with pre-ovulatory acquisition of developmental competence in bovine oocytes

Project description:This study investigated the cumulus cell (CC) transcriptomic changes during the oocyte developmental competence acquisition period. Six dairy cows were used for 24 oocyte collections and received FSH twice daily over 3 days, followed by FSH withdrawal for 20, 44, 68 and 92 h in four different oestrous cycles for each of the six cows. Half of the cumulusâoocyte complexes were subjected to in vitro maturation, fertilisation and culture to assess blastocyst rate. The other half of the CC underwent microarray analysis and qRT-PCR.

2016-04-01 | E-GEOD-79760 | biostudies-arrayexpress

Gene expression analysis of bovine oocytes with high developmental competence obtained from FSH-stimulated animals

Project description:The objective of the study was to analyze the impact of various lenght FSH withdrawal period (coasting) after the last FSH injection on transcriptome changes in in vivo bovine oocytes at the GV stage. Oocytes were collected from super-stimulated animals after 20, 44, 68 and 92 hours of coasting.

2013-06-21 | GSE38345 | GEO

Transcriptome analysis of FSH response in bovine cumulus cells

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2015-07-01 | E-GEOD-67970 | biostudies-arrayexpress

Transcriptome analysis of granulosa cells after conventional vs long FSH-induced superstimulation in cattle

Project description:Conventional Superstimulation (group 1) vs. Long Superstimulation (group 3) A genome-wide bovine oligo-microarray was used to compare the gene expression of granulosa cells collected from ovarian follicles after differing durations of the growing phase induced by exogenous FSH treatment. Cows were given a conventional (4-day) or long (7-day) superstimulatory treatment (25mg FSH im at 12-h intervals; n=6 per group), followed by prostaglandin treatment with last FSH and LH treatment 24 hr later. Granulosa cells were harvested 24 h after LH treatment. The expression of 416 genes was down-regulated and 615 genes was up-regulated in the long FSH group compared to the conventional FSH group. Quantification by RT-PCR of 7 genes (NTS, PTGS2, PTX3, RGS2, INHBA, CCND2 and LRP8) followed the same trend as in microarrays. Compared to the conventional group, the long FSH group responded to exogenous LH with down-regulation of mRNA for cyclinD2, lipoprotein receptorP8, CYP51A1, CYP19A1, CJA1, INHBA, SRPINE1, and up-regulation of Vannin, POSTN, GTPAse, Cysteine. Markers of fertility and follicle maturity were up-regulated in the long FSH group. We conclude that a prolonged FSH-induced growing phase is associated with transcriptomic characteristics of greater follicular maturity and may therefore be more appropriate for optimizing the superovulatory response and developmental competence of oocytes in cattle. straight comparison of Short Superstimulation group (the reference; group 1) versus a Long Superstimulation protocol ( the treatment; group 3) using 3 different animals (biological replicates) in each group and performed dye swap. For example on array 1: group 1 cow 1 versus group 3 cow 1 (cy3 vs cy5) and on array 4 is the dye swap group 3 cow 1 versus group 1 cow 1 (cy3 vs cy5).

2016-08-01 | E-GEOD-80289 | biostudies-arrayexpress

Impact of G protein versus beta-arrestin-dependent signal transduction at the FSH receptor (affy_fsh_human)

Project description:affy_fsh_human - affy_fsh_human - - G protein-coupled receptors (GPCR) are centrally involved in most physiological processes and are a major drug targets. They transduce extracellular signals inside the cells through at least two different mechanisms: i) the classical coupling to heterotrimeric G proteins and ii) a newly discovered beta-arrestin-dependent pathway. The fundamental issue of the respective impacts that these two transduction mechanisms exert on gene regulation has not been clearly addressed to date. To tackle this question, we have developed two mutants of the follicle stimulating hormone (FSH) receptors which do not couple to G proteins upon FSH activation but continue to recruit beta-arrestins and signal through them.-In the present study, we compare the wild-type FSH receptor to either the R466A or the T469F mutants. These two mutations are localized in the second intra cellular loop of the FSH receptor and prevent G protein coupling to the active FSH receptor. Each receptor was permanently expressed in HEK-293 cells at comparable levels. Cells were treated or not for 6 hours with 3 nM FSH. Keywords: treated vs untreated comparison,wt vs mutant comparison 18 arrays - Human GenomeU133A 2.0

2011-11-25 | E-GEOD-21982 | biostudies-arrayexpress

Gene expression analysis of bovine oocytes at optimal coasting time combined with GnRH antagonist during the no-FSH period

Project description:The objective of the study was to analyze the effect of a GnRH antagonist used during the FSH withdrawal period (coasting) on transcriptome changes in in vivo bovine oocytes at the GV stage. Oocytes were collected from super-stimulated animals after a coasting duration of 68hrs (optimal condition, control) and also from the same animal who received the GnRH antagonist (treated) during the whole coasting period. 68hrs of coasting (control) was compared with the 68hrs + GnRH antagonist (treatment), for each cow individually. Overall, 6 hybridizations were done, corresponding to the three biological replicates and one comparison using a dye-swap set-up.

2014-02-25 | E-GEOD-44664 | biostudies-arrayexpress

Chromatin maps of the Drosophila melanogaster TrxG protein Ash1 and FSH

Project description:Trithorax group (TrxG) proteins counteract Polycomb silencing by an as yet uncharacterized mechanism. A well-known member of the TrxG is the histone methyltransferase Absent, Small, or Homeotic discs 1 (ASH1). In Drosophila ASH1 is needed for the maintenance of Hox gene expression throughout development, which is tightly coupled to preservation of cell identity. In order to understand the molecular function of ASH1 in this process, we performed affinity purification of tandem-tagged ASH1 followed by mass spectrometry (AP-MS) and identified FSH, another member of the TrxG as interaction partner. Here we provide genome-wide chromatin maps of both proteins based on ChIP-seq. Our Dataset comprises of 4 ChIP-seq samples using chromatin from S2 cells which was immunoprecipitated, using antibodies against Ash1, FSH-L and FSH-SL.

2013-06-30 | E-GEOD-30820 | biostudies-arrayexpress

Expression data from mouse cumulus cells

Project description:Follicle stimulating hormone (FSH) and epidermal growth factor (EGF) are currently used on cumulus-oocyte complexes to mimic the luteinizing hormone surge in vitro and induce oocyte maturation and cumulus expansion. We have previously shown that addition of exogenous recombinant growth differentiation factor 9 (GDF9) during oocyte in vitro maturation led to an improvement of oocyte quality, as shown by an increased blastocyst percentage and fetal survival. Our objective was to characterize the effect of FSH/EGF and GDF9 treatments on mouse cumulus cells expression profile by microarray analysis. Cumulus-oocyte complexes (COCs) were recovered from 21 to 26 day old female 129/SV mice, 44 hours post equine chorionic gonadotropin treatment (eCG (5 IU)). For the microarray experiment whole COCs were treated with 293H control medium (0.125% v/v), with 20 ng/ml GDF9 or with a combination of 50 mIU/ml FSH and 10 ng/ml EGF. After 8 hours of in vitro maturation, COCs were denuded by gentle pipetting, the oocytes were removed and the cumulus cells centrifuged and extracted RNA analysed by microarray.

2011-12-10 | E-GEOD-19327 | biostudies-arrayexpress

Generation of chromatin maps for ASH1 and FSH applying ChIP-seq

Project description:see Super Series Summary Cross-linked chromatin derived from Drosophila S2-DRSC cells was immunoprecipitated using antibodies targeting ASH1 and FSH. Precipitated chromatin was sequenced applying Illumina sequencing.

2013-02-06 | E-GEOD-36404 | biostudies-arrayexpress

The BET protein FSH functionally interacts with ASH1 to orchestrate global gene activity in Drosophila

Project description:Background: The question of how cells re-establish gene expression states after cell division is still poorly understood. Genetic and molecular analyses have indicated that Trithorax group (TrxG) proteins are critical for the long-term maintenance of active gene expression states in many organisms. A generally accepted model suggests that TrxG proteins contribute to maintenance of transcription by protecting genes from inappropriate Polycomb group (PcG)-mediated silencing, instead of directly promoting transcription. Results: Here we report a physical and functional interaction in Drosophila between two members of the TrxG, the histone methyltransferase ASH1 and the bromodomain and extraterminal family protein FSH. We investigated this interface at the genome level, uncovering a widespread colocalization of both proteins at promoters and PcG-bound intergenic elements. Our integrative analysis of chromatin maps and gene expression profiles revealed that the observed ASH1-FSH binding pattern at promoters is a hallmark of active genes. Inhibition of FSH-binding to chromatin resulted in global down-regulation of transcription. In addition, we found that genes displaying marks of robust PcG-mediated repression also have ASH1 and FSH bound to their promoters. Conclusions: Our data strongly favor a global coactivator function of ASH1 and FSH during transcription, as opposed to the notion that TrxG proteins impede inappropriate PcG-mediated silencing, but are dispensable elsewhere. Instead, our results suggest that PcG repression needs to overcome the transcription-promoting function of ASH1 and FSH in order to silence genes. Refer to individual Series

2013-02-06 | E-GEOD-36450 | biostudies-arrayexpress

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