Effect of FSH on the gene expression profile of cumulus cells in vitro cultured as cumulus-oocyte complexes in presence of adenosine for 6 h
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ABSTRACT:
ORGANISM(S): Bos indicus Bos taurus
PROVIDER: GSE119803 | GEO | 2018/09/12
REPOSITORIES: GEO
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Project description:Changes in gene expression induced by adenosine treatment in cumulus cells using Agilent EmbryoGENE microarray slides. Bovine cumulus-oocyte complexes in vitro-cultured for 6 h using adenosine and compared to non-treated (control). After culture, COC were were washed and cumulus cells were used for microarray analysis
2018-09-12 | GSE119804 | GEO
Project description:Study the effect of recombinant human FSH supplementation in the in vitro maturation medium for an incubation of 6 hrs on the transcriptomics of bovine cumulus cells. 3 biological replicates of cumulus cells derived from cumulus oocyte complexes (COCs) supplemented with FSH (CC-FSH) vs. cumulus cells without FSH (Ct-6H, REFERENCE) were contrasted. Each replicate was subjected to a dye-swap procedure.
2015-07-01 | E-GEOD-67970 | biostudies-arrayexpress
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
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.
2011-12-10 | GSE19327 | GEO
Project description:To evaluate the mechanisms by which excessive FSH doses during ovarian stimulation are detrimental to ovarian follicle function and assisted reproduction outcomes, small ovarian reserve (SOR) heifers were subjected to ovarian stimulation with either a standard, 70 IU, or excessive, 210 IU, dose of commercial porcine FSH (cpFSH) preparation, per injection. The ovaries were recovered at the end of the ovarian stimulation protocol, consisting of eight cpFSH injections at 12 h intervals. In excessive dose treated heifers, heterogeneity was observed with regards to follicle phenotype including cumulus cell-oocyte complex (COC) morphology and intrafollicular steroid hormone milieu. In contrast, standard-dose treated heifers exhibited one predominant follicle phenotype which was selected as the control follicle type for all comparisons herein. RNA-seq analyses of granulosa, cumulus cells and the oocyte from individual follicles of these defined phenotypes was used to identify changes in the transcriptomes of these cell types in excessive dose treated individuals.
2022-08-10 | GSE197116 | GEO
Project description:impact of suppressing LH using the GnRH antagonist Cetrorelix during an ovarian coasting stimulation protocol on the transcriptome of bovine cumulus cells (CC). Oocytes were collected twice from six dairy cows. For the first collection, the cows received FSH twice daily for three days, followed by FSH withdrawal for 68 hours as control protocol. For the second collection, the same stimulation protocol was used, but the cows also received, starting on day 2 of FSH stimulation, a GnRH antagonist once a day until recovery of the cumulus-oocyte complexes (COC). Half of the COC were subjected to in vitro maturation, fertilization and culture to assess blastocyst rates. The other half of the CC underwent microarray analysis (n=3 cows, 2 treatments, 6 oocyte collections) and qRT-PCR (n=6 cows: 3 microarray cows + 3 other cows, 2 treatments, 12 oocyte collections). The differential expression of specific genes was confirmed by RT-qPCR: ATP6AP2, SC4MOL, OSTC and PTGDS. The global transcriptomic analysis of cumulus cells demonstrated that the inhibition of LH secretion may decrease survival, and growth of the follicle. Moreover, the results suggested that LH may be important to cumulus for the maintenance of cellular mechanisms such as global RNA expression, protein and nucleic acid metabolism and energy production. These results support the hypothesis that LH support is important during the final part of follicle maturation through its influence on the cumulus cells.
2018-10-26 | GSE121783 | GEO
Project description:Cumulus cells surrounding the oocyte were sampled at the following stages: developmentally incompetent or poorly competent prophase I oocytes (NC1 oocytes), developmentally competent prophase I oocytes (C1 oocytes), and developmentally competent metaphase II oocytes (C2 oocytes). NC1 samples were collected from immature, unexpanded cumulus-oocytes complexes (COC) from prepubertal (3-week-old) mice, C1 samples from immature, unexpanded cumulus-oocytes complexes (COC) from adult (8-week-old) and C2 samples from mature, expanded COCs obtained from the oviduct from 8-week-old mice after standard superovulation protocol.
2012-10-14 | GSE36604 | GEO
Project description:Cumulus cells surrounding the oocyte were sampled at the following stages: developmentally incompetent or poorly competent prophase I oocytes (NC1 oocytes), developmentally competent prophase I oocytes (C1 oocytes), and developmentally competent metaphase II oocytes (C2 oocytes). NC1 samples were collected from immature, unexpanded cumulus-oocytes complexes (COC) from prepubertal (3-week-old) mice, C1 samples from immature, unexpanded cumulus-oocytes complexes (COC) from adult (8-week-old) and C2 samples from mature, expanded COCs obtained from the oviduct from 8-week-old mice after standard superovulation protocol. Global transcriptional profiling was performed using cumulus cells collected from murine ovarian follicles during in vivo oocyte developmental competence acquisition. Cumulus cells were collected at 3 stages: early stage follicles (prophase I arrested oocytes, meiotically competent but developmentally incompetent, n=5), late stage follicles (prophase I arrested oocytes, meiotically competent and developmentally competent, n=5) and ovulatory follicles collected in vivo (metaphase II arrested oocytes, developmentally fully competent, n=5).
2012-10-14 | E-GEOD-36604 | biostudies-arrayexpress
Project description:Ovaries from cyclic sows were derived from a local slaughterhouse and transported to the laboratory at 27-33ºC within 2 hours after death. After removal of surrounding tissue, the ovaries were rinsed with water (27-33ºC) and submersed in PBS containing 100U/ml penicillin and 100µg/ml streptomycin (Gibco, Breda, The Netherlands) at 37ºC. Granulosa cells and oocytes were aspirated with a 26G needle from healthy, 3-6 mm follicles. Follicular health was determined according to previously described criteria (28). Harvested cells were pooled and the cell suspension was washed twice by centrifugation at room temperature in McCoys 5a medium with glutamine (Invitrogen, Breda, The Netherlands) supplemented with 100U/ml penicillin, 100µg/ml streptomycin, 0.1% BSA (Fluka, Zwijndrecht, The Netherlands), 2.5µg/ml transferrin and 4ng/ml sodium selenite. Oocytes were removed from the cell suspension by filtration through a 70µm cell sieve (Becton en Dickinson, Alphen a/d Rijn, The Netherlands). The number of viable GCs was estimated by trypan blue exclusion. Granulosa cells were cultured in serum free medium as previously described by Picton and colleagues (26). For co-culture experiments cumulus oocyte complexes (COC) were aspirated from 3-6mm follicles. Cell debris and COC were allowed to settle at the bottom of the tube and the supernatant was discarded. Cumulus oocyte complexes were washed in TL-HEPES and subsequently selected by morphological appearance (dark cytoplasm and at least 2 layers of cumulus cells). The COC were transferred to wash medium containing 0.1U/l FSH, 100mg/l long R3-IGF-1, 10mg/l insulin (all from Sigma Aldrich, Zwijndrecht, The Netherlands) and 100mg/l testosterone (Fluka, Zwijndrecht, The Netherlands). Granulosa cells, 7.5*106 per well of a 6 wells plate, and COC, 80-100, were co-cultured in 3.5ml medium per well of a 6 wells plate. Cells were cultured in a humidified atmosphere at 37ºC and 5% CO2 for 48 hrs.
2010-03-31 | E-MEXP-1768 | biostudies-arrayexpress
Project description:In cultured cumulus oocyte complexes (COC) FSH induces gene transcription required for germinal vesicle breakdown (GVBD). Experiments were performed to determine the critical period when gene transcription is required for GVBD and identify candidate mRNAs involved. Experiment I: murine COC were cultured 4h in the presence of FSH with 5,6-dichloro-1-beta-D-ribofuranosylbenzimidazole (DRB) added at different intervals after the start of culture. COC cultured with FSH underwent GVBD (82+/-7%). When DRB was added at 0, 5 or 10 min after culture initiation, oocyte maturation was blocked (17+/-7%, 14+/-6% and 21+/-6% GVBD, respectively). When DRB was added after 15, 20 or 30 min, progressively more COC underwent GVBD (37+/-6%, 39+/-6% and 66+/-6%, respectively). The critical period of transcription required for GVBD occurred between 15 and 30 min after culture initiation. Experiment II: COC were cultured for 25 min in the presence (plusDRB) or absence (minusDRB) of DRB. SAGE libraries were generated from COC RNA of each treatment group. A total of 48,431 and 45,367 tags were sequenced for the plusDRB and minusDRB libraries, respectively. Criteria used to identify transcripts of interest included a total tag count of at least 10 across both libraries and a 3-fold or greater difference in expression between libraries. Using these criteria, 39 and 27 transcripts were identified as differentially expressed at the P</=0.01 and P</=0.001 levels, respectively. Differentially expressed transcripts were classed into major categories that included cell growth, development and regulation of gene expression. Differentially expressed transcripts represent candidates potentially involved in regulating maturation of murine COC. Keywords: Analysis of treatment effect using SAGE library comparisons Mouse cumulus oocyte complexes (COC) cultured 25 min in presence (plusDRB) or absence (minusDRB) of 120 uM DRB, a transcriptional inhibitor. SAGE libraries were constructed from 9 to 10 ug RNA from each treatment group. Approx. 48,000 tags seqenced from each library. Comparison of these 2 libraries used to identify potential candidate mRNAs associated with regulation of germinal vesicle breakdown in murine COC.
2006-08-31 | E-GEOD-5568 | biostudies-arrayexpress