Gene expression analysis of granulosa cells from Fzd1 WT and KO mice
Ontology highlight
ABSTRACT:
ORGANISM(S): Mus musculus
PROVIDER: GSE38315 | GEO | 2012/09/01
SECONDARY ACCESSION(S): PRJNA167694
REPOSITORIES: GEO
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Project description:WNT4 is required for normal ovarian follicle development and female fertility in mice, but how its signal is transduced remains unknown. Fzd1 encodes a WNT receptor whose expression is markedly induced in both mural granulosa cells and cumulus cells during the preovulatory period, in a manner similar to Wnt4. To study the physiological roles of FZD1 in ovarian physiology and to determine if it serves as receptor for WNT4, Fzd1-null mice were created by gene targeting. Whereas rare Fzd1-/- females were sterile due to uterine fibrosis and ovarian tubulostromal hyperplasia, the majority were subfertile, producing M-bM-^IM-^H1 less pup per litter on average relative to controls. Unlike WNT4-deficient mice, ovaries from Fzd1-/- mice had normal weights, numbers of follicles, steroid hormone production and WNT4 target gene expression levels. Microarray analyses of granulosa cells from periovulatory follicles revealed few genes whose expression was altered in Fzd1-/- mice. However, gene expression analyses of cumulus-oocyte complexes (COCs) revealed a blunted response of both oocyte (Zp3, Dppa3, Nlrp5, Bmp15) and cumulus (Btc, Ptgs2, Sema3a, Ptx3, Il6, Nts, Alcam, Cspg2) genes to the ovulatory signal, whereas the expression of these genes was not altered in WNT4-deficient COCs from Wnt4tm1.1Boer/tm1.1Boer;Tg(CYP19A1-cre)1Jri mice. Despite altered gene expression, cumulus expansion appeared normal in Fzd1-/- COCs both in vitro and in vivo. Together, these results indicate that Fzd1 is required for normal female fertility and may act in part to regulate oocyte maturation and cumulus cell function, but is unlikely to function as the sole ovarian WNT4 receptor. Triplicate RNA samples from granulosa cells of Fzd1 KO mice are compared to triplicate RNA samples from granulosa cells of control Fzd1 WT mice
2012-09-01 | E-GEOD-38315 | biostudies-arrayexpress
Project description:We are human embryologists from center for reproductive medicinel of Anhui Provincial Hospital Affiliated to Anhui Medical University, and we have the expertise to do all that properly in humans. By deep sequencing, the current experiment determined the miRNA profile of two intrafollicular somatic cell types: CRCs and COCs, isolated from women undergoing controlled ovarian stimulation and in vitro fertilization treatment. Ovarian follicles, which are a densely-packed shell of granulosa cells that contains an immature or mature oocyte, are above all responsible for the development, maturation, and release of mature egg for fertilization. They are also responsible for synthesizing and secreting hormones that are essential for follicular development, menstrual and estrous cycle, maintenance of the reproductive tracts and their functions, development of female secondary sex characteristics, and metabolism. During folliculogenesis, ovarian granulosa cells surrounding the oocyte differentiate into mural granulosa cells, involved in gonadal steroidogenesis, and into cumulus cells, which are ovulated with the oocyte at ovulation. These cumulus cells derive from the same population of early follicles, but differentiate into two distinct groups of cells: 1) Those directly lie on the zona pellucida are composed of the so called corona radiata cells.(CRCs) 2) The other group surrounds the CRCs and consists of more numerous cells, forming the so called cumulus oophorus cells (COCs). In the present study, we described the miRNA expression profile to characterize the ensemble of both known and novel miRNAs expressed in CRCs, as well as in COCs, by using high-throughput Solexa technology.
2014-07-14 | E-MTAB-2264 | biostudies-arrayexpress
Project description:Ovulation requires sequential molecular events and structural remodeling in the ovarian follicle for the successful release of a mature oocyte capable of being fertilised. Critical to this process is progesterone receptor (PGR), a transcription factor highly yet transiently expressed in granulosa cells of preovulatory follicles. Progesterone receptor knockout (PRKO) mice are anovulatory, with a specific and complete defect in follicle rupture. Therefore, this model was used to examine the critical molecular and biochemical mechanisms necessary for successful ovulation. Although PGR is not expressed in the cumulus cells or oocyte of the preovulatory cumulus oocyte complex (COC), it is well known that the COC responds to the cascade of gene expression changes that occurs in preovulatory granulosa cells. We used microarrays to identify putative ‘ovulation’ genes in preovulatory COCs at a time when PGR expression is maximal in granulosa cells (eCG + 8h hCG) and the preovulatory COC and follicle are undergoing the final changes necessary for successful ovulation.
2018-04-20 | GSE92436 | 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:To study the physiological role of WNT4 in the postnatal ovary, a mouse strain bearing a floxed Wnt4 allele was created and mated to the Amhr2tm3(cre)Bhr strain to target deletion of Wnt4 to granulosa cells. Wnt4flox/-;Amhr2tm3(cre)Bhr/+ mice had significantly reduced ovary weights and produced smaller litters (P<0.05). Serial follicle counting demonstrated that, while Wnt4flox/-;Amhr2tm3(cre)Bhr/+ mice were born with a normal ovarian reserve and maintained normal numbers of small follicles until puberty, they had only 25.2% of the normal number of healthy antral follicles. Some Wnt4flox/-;Amhr2tm3(cre)Bhr/+ mice had no antral follicles or corpora lutea and underwent premature follicle depletion. RTPCR analyses of Wnt4flox/-;Amhr2tm3(cre)Bhr/+ granulosa cells and cultured granulosa cells that overexpress WNT4 demonstrated that WNT4 regulates the expression of Star, Cyp11a1 and Cyp19, steroidogenic genes previously identified as downstream targets of the WNT signaling effector CTNNB1. WNT4- and CTNNB1-overexpressing cultured granulosa cells were analyzed by microarray for alterations in gene expression, which showed that WNT4 also regulates a series of genes involved in late follicle development and the cellular stress response via the WNT/CTNNB1 signaling pathway. Together, these data indicate that WNT4 is required for normal antral follicle development, and may act by regulating granulosa cell functions including steroidogenesis.
2009-10-23 | GSE18704 | GEO
Project description:To study the physiological role of WNT4 in the postnatal ovary, a mouse strain bearing a floxed Wnt4 allele was created and mated to the Amhr2tm3(cre)Bhr strain to target deletion of Wnt4 to granulosa cells. Wnt4flox/-;Amhr2tm3(cre)Bhr/+ mice had significantly reduced ovary weights and produced smaller litters (P<0.05). Serial follicle counting demonstrated that, while Wnt4flox/-;Amhr2tm3(cre)Bhr/+ mice were born with a normal ovarian reserve and maintained normal numbers of small follicles until puberty, they had only 25.2% of the normal number of healthy antral follicles. Some Wnt4flox/-;Amhr2tm3(cre)Bhr/+ mice had no antral follicles or corpora lutea and underwent premature follicle depletion. RTPCR analyses of Wnt4flox/-;Amhr2tm3(cre)Bhr/+ granulosa cells and cultured granulosa cells that overexpress WNT4 demonstrated that WNT4 regulates the expression of Star, Cyp11a1 and Cyp19, steroidogenic genes previously identified as downstream targets of the WNT signaling effector CTNNB1. WNT4- and CTNNB1-overexpressing cultured granulosa cells were analyzed by microarray for alterations in gene expression, which showed that WNT4 also regulates a series of genes involved in late follicle development and the cellular stress response via the WNT/CTNNB1 signaling pathway. Together, these data indicate that WNT4 is required for normal antral follicle development, and may act by regulating granulosa cell functions including steroidogenesis. Experiment Overall Design: Granulosa cells were obtained from 20-26 day-old mice of various genotypes 48h after eCG treatment, using the needle puncture method as previously described. Cells were then infected with adenoviruses to express eGFP or Cre, or overexpress WNT4 in serum-free medium, and subsequently harvested for RNA extraction as described below. Preliminary experiments demonstrated that an infection efficiency of >80% could be obtained at an MOI of ~50 (as determined by analysis of fluorescent signal in Ad-eGFP-infected cells) and that the Ad-Cre and Ad-WNT4 viruses produced efficient recombination of the floxed Ctnnb1 alleles and robust WNT4 overexpression, respectively. Microarray analyses were done using triplicate RNA samples from each adenoviral treatment described above, and using mouse expression set 430 microarrays (Affymetrix, Santa Clara, CA).
2009-11-07 | E-GEOD-18704 | biostudies-arrayexpress
Project description:This group consist of human embryologists from the reproductive medical center for of the 1st affiliated hospital of Anhui Medical University. Our research is specifically focused on women ovarian reserve and the relevant female infertility. By deep sequencing, the current experiment determined the small non-coding RNA profile of cumulus cells from patients with or without diminished ovarian reserve undergoing controlled ovarian stimulation and in vitro fertilization treatment. Ovarian follicles, which are a densely-packed shell of granulosa cells that contains an immature or mature oocyte, are above all responsible for the development, maturation, and release of mature egg for fertilization. They are also responsible for synthesizing and secreting hormones that are essential for follicular development, menstrual and estrous cycle, maintenance of the reproductive tracts and their functions, development of female secondary sex characteristics, and metabolism. During folliculogenesis, ovarian granulosa cells surrounding the oocyte differentiate into mural granulosa cells, involved in gonadal steroidogenesis, and into cumulus cells, which are ovulated with the oocyte at ovulation. In the present study, we described the small non-coding RNA expression profile to characterize the ensemble of both known and novel ncRNAs expressed in cumulus cells from patients with or without Diminished ovarian reserve, by using high-throughput Solexa technology.
2015-11-20 | E-MTAB-3913 | biostudies-arrayexpress
Project description:This SuperSeries is composed of the following subset Series: GSE36602: Transcriptional profiling of somatic cells differentiation throughout oocyte competence acquisition in rainbow trout ovarian follicles. GSE36603: Transcriptional profiling of somatic cells differentiation throughout oocyte competence acquisition in the xenopus ovarian follicles. GSE36604: Transcriptional profiling of cumulus cells differentiation throughout oocyte competence acquisition in the murine ovarian follicles. GSE36605: Transcriptional profiling of cumulus cells differentiation throughout oocyte competence acquisition in the bovine ovarian follicles. Refer to individual Series
2012-10-14 | E-GEOD-36617 | biostudies-arrayexpress
Project description:The antral follicle stage plays a crucial role in mammalian oocyte maturation, signifying the final stages of oocyte development and ovulation. This complex process relies on synchronized interactions between oocyte maturation and the proliferation of neighboring granulosa cells. Previous studies have identified two subtypes of granulosa cells in antral follicles: cumulus granulosa cells, located in the inner region, which surround and support the oocyte, and mural granulosa cells, present in the outer layers, which provide mechanical support to the follicular wall and possess steroidogenic functions. Despite the wealth of molecular data generated by these studies, many fundamental questions regarding key developmental events, granulosa cell heterogeneity, functional annotation, and the intricate relationship between somatic cells and oocytes still lack detailed, single-cell resolution-level investigations. In this study, we isolated follicular cells from porcine antral follicles and conducted scRNA-seq to analyze the single-cell transcriptomes of these cells. By identifying and sub-clustering ovarian cells, such as mural granulosa cells and cumulus granulosa cells, we elucidated the heterogeneity of granulosa cells in pigs.
2023-10-24 | GSE237134 | GEO
Project description:Despite great improvements in assisted reproductive technology (ARTs), the success of in vitro embryo production remains relatively low. Most of the oocytes used to produce in vitro embryos are recovered from smaller follicles, forming a heterogeneous population that must be matured in vitro. Since the original in vitro maturation (IVM ) experiments (Heilbrunn et al. , 1939) the process by which the most competent oocytes are selected to produce blastocysts remains similar and is still based on morphological aspects of the oocyte cytoplasm and the number of layers and compaction of cumulus cells attached to the oocyte surface (Armstrong, 2001, Coticchio et al. , 2004, Krisher, 2003, Lonergan et al. , 2003). Ovaries from crossbred cows (Bos indicus x Bos taurus) were collected immediately after slaughter and transported to the laboratory in saline solution (0.9% NaCl) supplemented with penicillin G (100 IU/mL) and streptomycin sulfate (100 mg/mL) at 35-37C. The follicles were dissected from the ovarian cortex using scissors, scalpels and tweezers in TCM-199 medium supplemented with Hank’s salts and 10% fetal calf serum (FCS; GIBCO BRL) at 36C. Follicles were measured using a graduated eyepiece (OSM-4; Olympus) and then classified morphologically into two groups according to their diameter: 1.0-3.0 mm or ≥8.0 mm. The criteria used for follicle selection included the presence of extensive and fine vascularization and a shiny and translucent appearance. After follicular rupture, the presence of granulosa cells with a regular and healthy appearance and no free-floating particles in the follicular fluid were also used as selection criteria. Only COCs with a homogeneous granulated cytoplasm and at least three layers of compact of cumulus cells were used in the present study. The selected COCs were transferred to a 50 μL droplet of phosphate-buffered saline (PBS), and the cumulus cells were mechanically removed by repeated pipetting. After cumulus collection, they were transferred to a 0.2-mL tube and centrifuged twice for 2 min at 700 g. The supernatant was removed, and 2μL of RNAlater (Applied Biosystems, Foster City, CA, USA) was added to the pellet, which was then stored at -20C until RNA extraction. For each follicle size group, three replicas of pooled cumulus cells corresponding to 30 oocytes were stored for RNA extraction and subsequent microarray assays, and four independent replicates corresponding to 20 oocytes were stored for RNA extraction for subsequent qPCR assays. During oogenesis, the somatic cells that surround the oocyte proliferate and differentiate into cumulus cells (CCs), which are metabolically coupled to the oocyte, forming the cumulus-oocyte complexes (COCs) (van den Hurk and Zhao, 2005). The CCs remain in strong contact with the oocyte, maintained by cytoplasmic bridges called gap-junctions, as well as through justacrine and paracrine signaling networks (Albertini et al. , 2001, Gilchrist et al. , 2004, Tanghe et al. , 2002, Vozzi et al. , 2001, Webb et al. , 2002). This intense bidirectional communication between the CCs and the oocyte is maintained during all phases of folliculogenesis and is essential for the acquisition of oocyte competence that is required for blastocyst development (Assidi et al., 2008, Fair, 2003, Gilchrist et al., 2004). An informative model to access the level of oocyte competence that has been used by many research groups is follicle size (Donnison and Pfeffer, 2004, Franco et al. , 2013, Lequarre et al. , 2005, Mourot et al., 2006). In a previous study, we showed that oocytes obtained from follicles 1-3 mm in diameter are significantly less competent in producing blastocysts than oocytes obtained from follicles ≥8 mm in diameter (Franco et al., 2013). In the present study, we use the same model to compare the gene expression profile of more than 23,000 bovine transcripts between the CCs obtained from incompetent COCs (1-3 mm) and competent COCs (≥8 mm). We have found substantial differences in the expression of several gene clusters representing distinct metabolic pathways such as energy metabolism, cell signaling, cell cycle, DNA repair, meiosis, and inflammation.
2015-01-27 | E-GEOD-65269 | biostudies-arrayexpress