Transcription profiling by array of granulosa and cumulus cells following ovulation induction with either hCG or a GnRH agonist trigger to explore differences in follicle transcriptomes in patients
ABSTRACT: The objective of this experiment was to explore differences in follicle transcriptomes in patients having oocyte maturation induced with either a bolus of hCG or a GnRH agonist. Upon oocyte retrieval, mural granuloasa cells and cumulus cells were isolated, allowing for comparison of the follicular transcriptomes between patients treated with eihter hCG or the GnHR agonist.
Project description:Comparison of 9 paired human granulosa cell samples just before and 36h after hCG triggering for controlled ovarian stimulation, to explore genes regulated by hCG and involved with ovulation and final oocyte maturation.
Project description:Assisted reproduction technologies (ART) and high selection pressure observed in the dairy industry are leading towards the use of younger females for reproduction purpose, reducing the interval between generations. This situation might have an impact on 10 young Holstein cows were used 3 times each at different ages for ovarian stimulation protocols and oocyte collections (at 8, 11 and 14 months). These oocytes were then fertilized in vitro with adult bull semen, generating 3 lots of embryos per animal.
Project description:Polycystic ovary syndrome (PCOS) is a common endocrine and metabolic disorder that is characterized by increased circulating androgen levels, anovulatory infertility, and frequently, insulin resistance and hyperinsulinemia.The abnormity of oocyte nuclear maturity is the main reason for anovulatory infertility and pregnancy loss in PCOS patients.The bidirectional exchanges between oocyte and contiguous CCs are important for oocyte competence acquisition, early embryonic development and CC expansion.Gene expression profiles of CCs has been suggested to predict embryo development and pregnancy outcome. We used microarrays to detail the global programme of gene expression of CCs isolated from oocytes at metaphase I (CCMI) and metaphase II (CCMII) stage under controlled ovarian stimulation (COS) cycle in PCOS patients. Cumulus cells were isolated from oocyte at stage metaphase 1(MI)and stage metaphase II (MII) of PCOS patients for RNA extraction and hybridization on Affymetrix microarrays. For microarray analysis, we used three chips for each CC category. That is, CCMI1,CCMI2,CCMI3,CCMII1,CCMII2 and CCMII3.
Project description:Influence of ovarian stimulation with 200 IU of hCG, (administered in the late follicular phase among ICSI patients undergoing a GnRH-antagonist protocol), on the endometrium on the day of oocyte pick-up. The purpose of the present study is to assess the influence of the administration of low dose hCG on the endometrium. In addition, by analysing the correlation of the morphological pattern and gene expression profile of human endometrium on the day of oocyte retrieval in patients of both treatment groups, we want to study the implantation potential. Pregnant and non-pregnant patients were compared. In total 6 samples were analyzed for gene expression analysis with microarrays, 3 pregnant and 3 non-pregnant patients were compared.
Project description:The aim of the study was to describe and characterise the phosphodiesterases in the human ovary. Part of the study included results from analysis of mRNA microarray data from follicles and granulosa cells from three previously published studies(E-MEXP-3783, E-MTAB-2203, E-MTAB-1670). In addition, we used data from two unpublished studies with granulosa cells isolated from 4-6 mm antral follicles and preantral follicles. The included studies covered the folliculogenesis from the preantral stage to after induction of ovulation. For comparison, previously published dataset from heart (E-GEOD-22253, E-MEXP-2654), parietal cortex (E-GEOD-35977), cerebellum (E-GEOD-35974), lung (E-GEOD-43458), and peripheral blood mononucleated cells(E-GEOD-23832) were included in addition to various tissues from Affymetrix's sample data set.
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).
Project description:Premature progesterone (P) rise during GnRH antagonist cycles for IVF is a frequent phenomenon and has been associated with lower pregnancy and implantation rates. Different thresholds of progesterone have been used so far to define its premature rise during the follicular phase of an IVF stimulated cycle. In this study, we evaluated endometrial gene expression on the day of oocyte retrieval according to the level of serum progesterone on the day of hCG administration in GnRH antagonist cycles.Endometrial biopsies from eleven patients were taken with a Pipelle de Cornier (Prodimed, Neuilly-en-Thelle, France) on the day of oocyte retrieval in a GnRH antagonist/rec-FSH stimulated IVF cycle with fresh embryo transfer. Biopsies were analysed for gene expression with Affymetrix Human Genome (HG) U133 Plus 2.0 Arrays and GCOS software (Affymetrix, Santa Clara, CA, USA). Patients were divided into three different groups according to their progesterone serum concentration on the day of hCG administration (A) P <= 0.9 ng/mL, (B) 1 < P < 1.5 ng/mL, and (C) P > 1.5 ng/mL. Serum P was measured with the automated Elecsys immunoanalyser (Roche Diagnostics, Mannheim, Germany). Selected differentially expressed genes were validated with quantitative real-time PCR (QPCR) with TaqMan Gene Expression Assays (Applied Biosystems, Foster City, CA, USA). Keywords: gene expression analysis, premature progesterone rise Endometrial biopsies from fourteen patients were taken on the day of oocyte retrieval in a GnRH/rec-FSH stimulated IVF cycle with fresh embryo transfer and analyzed with microarrays. Patients were divided into three groups according to their progesterone serum concentration on day of hCG: A <=0.9ng/ml; B 1-1.5 ng/ml; C > 1.5ng/ml
Project description:Assisted reproduction technologies (ART) and high selection pressure observed in the dairy industry are leading towards the use of younger females for reproduction purpose, reducing the interval between generations. This situation might have an impact on embryo quality, which can affect the success rate of the procedures. This study aims to document the effect of donor age on embryo quality, on transcriptomic level, in order to caracterize the effect of using young animals for reproduction purpose. 10 young Holstein cows were used 3 times each at different ages for ovarian stimulation protocols and oocyte collections (at 8, 11 and 14 months). These oocytes were then fertilized in vitro with adult bull semen, generating 3 lots of embryos per animal. Semen from the same bull was used for all females, at all times. Each animal was used as its own control for age-effect evaluation. EmbryoGENE plateform allowed whole genome assessment of gene expression patterns at the blastocyst stage. Embryos from animals at 8 vs 14 months and at 11 vs 14 months were used for microarray hybridization. Validation was done performing RT-qPCR tests on 7 candidate genes.
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.
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 cumulus–oocyte 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.