Transcriptomic profiling of bovine ovarian cortical tissue cultured with FSH
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ABSTRACT:
ORGANISM(S): Bos taurus
PROVIDER: GSE147176 | GEO | 2020/07/29
REPOSITORIES: GEO
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Project description:The forkhead box transcription factor FOXO1 is highly expressed in granulosa cells of growing follicles but is down-regulated by FSH in culture or by LH-induced luteinization in vivo. To analyze the function of FOXO1, we infected rat and mouse granulosa cells with adenoviral vectors expressing two FOXO1 mutants: a gain-of-function mutant FOXOA3 that has two serine residues and one threonine residue mutated to alanines rendering this protein constitutively active and nuclear, and a FOXOA3-mutant DNA-binding domain (mDBD) in which the DBD is mutated. The infected cells were then treated with vehicle or FSH for specific time intervals. Infection of the granulosa cells was highly efficient, caused only minimal apoptosis, and maintained FOXO1 protein at levels of the endogenous protein observed in cells before exposure to FSH. RNA was prepared from control and adenoviral infected cells exposed to vehicle or FSH for 12 and 24 h. Affymetrix microarray and database analyses identified, and real time RT-PCR verified, that genes within the lipid, sterol, and steroidogenic biosynthetic pathways (Hmgcs1, Hmgcr, Mvk, Sqle, Lss, Cyp51, Tm7sf2, Dhcr24 and Star, Cyp11a1, and Cyp19), including two key transcriptional regulators Srebf1 and Srebf2 of cholesterol biosynthesis and steroidogenesis (Nr5a1, Nr5a2), were major targets induced by FSH and suppressed by FOXOA3 and FOXOA3-mDBD in the cultured granulosa cells. By contrast, FOXOA3 and FOXOA3- mDBD induced expression of Cyp27a1 mRNA that encodes an enzyme involved in cholesterol catabolism to oxysterols. The genes up-regulated by FSH in cultured granulosa cells were also induced in granulosa cells of preovulatory follicles and corpora lutea collected from immature mice primed with FSH (equine choriogonadotropin) and LH (human choriogonadotropin), respectively. Conversely, Foxo1 and Cyp27a1 mRNAs were reduced by these same treatments. Collectively, these data provide novel evidence that FOXO1 may play a key role in granulosa cells to modulate lipid and sterol biosynthesis, thereby preventing elevated steroidogenesis during early stages of follicle development.
2009-04-17 | GSE15727 | GEO
Project description:The Pik3cd null females are subfertile and have less growing follicles than their heterozygous littermates in the ovary. These mice poorly respond to the exogenous gonadotropins and ovulate much less oocytes than controls. In addition, the estrodial stimulated GC proliferation in preantral follicles is also impaired in Pik3cd null ovaries. FSH and E2 dramatically activates PI3K/AKT pathway in GCs of wild type mice, but not in the Pik3cd null mice. we used microarray to identify those genes regulated by Pik3cd in response to PMSG (Pregnant Mare Serum Gonadotropin) and E2 treatment in ovary.
2012-10-23 | GSE41746 | GEO
Project description:The forkhead box transcription factor FOXO1 is highly expressed in granulosa cells of growing follicles but is down-regulated by FSH in culture or by LH-induced luteinization in vivo. To analyze the function of FOXO1, we infected rat and mouse granulosa cells with adenoviral vectors expressing two FOXO1 mutants: a gain-of-function mutant FOXOA3 that has two serine residues and one threonine residue mutated to alanines rendering this protein constitutively active and nuclear, and a FOXOA3-mutant DNA-binding domain (mDBD) in which the DBD is mutated. The infected cells were then treated with vehicle or FSH for specific time intervals. Infection of the granulosa cells was highly efficient, caused only minimal apoptosis, and maintained FOXO1 protein at levels of the endogenous protein observed in cells before exposure to FSH. RNA was prepared from control and adenoviral infected cells exposed to vehicle or FSH for 12 and 24 h. Affymetrix microarray and database analyses identified, and real time RT-PCR verified, that genes within the lipid, sterol, and steroidogenic biosynthetic pathways (Hmgcs1, Hmgcr, Mvk, Sqle, Lss, Cyp51, Tm7sf2, Dhcr24 and Star, Cyp11a1, and Cyp19), including two key transcriptional regulators Srebf1 and Srebf2 of cholesterol biosynthesis and steroidogenesis (Nr5a1, Nr5a2), were major targets induced by FSH and suppressed by FOXOA3 and FOXOA3-mDBD in the cultured granulosa cells. By contrast, FOXOA3 and FOXOA3- mDBD induced expression of Cyp27a1 mRNA that encodes an enzyme involved in cholesterol catabolism to oxysterols. The genes up-regulated by FSH in cultured granulosa cells were also induced in granulosa cells of preovulatory follicles and corpora lutea collected from immature mice primed with FSH (equine choriogonadotropin) and LH (human choriogonadotropin), respectively. Conversely, Foxo1 and Cyp27a1 mRNAs were reduced by these same treatments. Collectively, these data provide novel evidence that FOXO1 may play a key role in granulosa cells to modulate lipid and sterol biosynthesis, thereby preventing elevated steroidogenesis during early stages of follicle development. Experiment Overall Design: Immature rat granulosa cells were affected with either control (GFP) or FOXO1 mutant adenovirus (FOXOA3 and FOXOA3-mDBD) for 4 hours. Then the cells were treated with either FSH or vehicle for 24 hours. The gene expression profiles for the four treatment samples (control, control +FSH, FOXOA3 + FSH, FOXOA3-mDBD + FSH) were compared using Rat Genome 230.2 Arrays with one array per sample.
2009-04-25 | E-GEOD-15727 | biostudies-arrayexpress
Project description:To unravel the genes that underpin the effects of FSH on spermatogenesis, testicular gene expression was compared in juvenile rats after FSH suppression to littermate controls. Based on existing data, it is apparent that the dominant function of FSH shifts between 9 and 18 day postpartum (dpp) during the first wave of spermatogenesis from driving Sertoli cell proliferation to support of germ cells. To enable comprehensive analysis of the impact of acute in vivo FSH suppression on Sertoli and germ cell development and to determine the genes that underpin these affects, FSH was selectively suppressed in Sprague Dawley rats by passive immunisation for 4 days prior to testis collection at 18 days postpartum. FSH suppression did not affect Sertoli cell proliferation at 18 dpp, but germ cell numbers were decreased at 18 dpp following FSH suppression, with a corresponding increase in germ cell apoptosis measured. Sixty transcripts were measured as changed at 18 dpp in response to 4 days of FSH suppression, as assessed using Affymetrix™ microarrays. Some of these are known as Sertoli cell-derived FSH-responsive genes (e.g. StAR, cathepsin L, insulin-like growth factor binding protein-3), while others encode proteins involved in cell cycle and survival regulation (e.g. cyclin D1, scavenger receptor class B 1). These data demonstrate that FSH affects germ cells in vivo, by supporting germ cell viability at day 18. This model enabled identification of candidate genes that contribute to the FSH-mediated pathway by which Sertoli cells support germ cells. Keywords: time course
2008-01-29 | GSE10293 | GEO
Project description:Background: Secreted frizzled-related proteins (SFRPs) comprise a family of WNT signaling antagonists whose roles in the ovary are poorly understood. Sfrp4-null mice were previously found to be hyperfertile due to an enhanced granulosa cell response to gonadotropins, leading to decreased antral follicle atresia and enhanced ovulation rates. The present study aimed to elucidate the mechanisms whereby Sfrp4 antagonizes FSH action. Methods: Primary cultures of granulosa cells from wild-type mice were treated with FSH and/or SFRP4, and effects of treatment on gene expression were evaluated by RT-qPCR and RNAseq. Bioinformatic analyses were conducted to analyse the effects of SFRP4 on the transcriptome, and compare them to those of FSH or a constitutively active mutant of FOXO1. Additional granulosa cell cultures from wild-type or Sfrp4-null mice, some pretreated with pharmacologic inhibitors of specific signaling effectors, were used to examine the effects of FSH and/or SFRP4 on signaling pathways, autophagy and apoptosis by western blotting and TUNEL. Results: Treatment of cultured granulosa cells with recombinant SFRP4 was found to decrease basal and FSH-stimulated mRNA levels of FSH target genes. Unexpectedly, this effect was found to occur neither via a canonical (CTNNB1-dependent) nor non-canonical WNT signaling mechanism, but was found to be GSK3β-dependent. Rather, SFRP4 was found to antognize AKT activity via a mechanism involving AMPK. This lead to the hypophosphorylation of FOXO1 and a decrease in the expression of a portion of the FSH and FOXO1 transcriptomes. Conversely, FSH-stimulated AMPK, AKT and FOXO1 phosphorylation levels were found to be increased in the granulosa cells of Sfrp4-null mice relative to wild-type controls. SFRP4 treatement of granulosa cells also induced autophagy, both by signaling via AKT-mTORC1-ULK1 and by transcriptional regulation of autophagy-related genes. SFRP4 further induced granulosa cell apoptosis in an autophagy-independent manner. Conclusions: This study identifies a novel GSK3β-AMPK-AKT signaling mechanism through which SFPR4 antagonizes FSH action, and further identifies SFRP4 as a novel regulator of granulosa cell autophagy. These findings provide a mechanistic basis for the phenotypic changes previously observed in Sfrp4-null mice, and broaden our understanding of the physiological roles of WNT signaling processes in the ovary.
2024-08-21 | GSE266715 | GEO
Project description:The Pik3cd null females are subfertile and have less growing follicles than their heterozygous littermates in the ovary. These mice poorly respond to the exogenous gonadotropins and ovulate much less oocytes than controls. In addition, the estrodial stimulated GC proliferation in preantral follicles is also impaired in Pik3cd null ovaries. FSH and E2 dramatically activates PI3K/AKT pathway in GCs of wild type mice, but not in the Pik3cd null mice. we used microarray to identify those genes regulated by Pik3cd in response to PMSG (Pregnant Mare Serum Gonadotropin) and E2 treatment in ovary. We set 3 treatment groups of both Pik3cd+/- and Pik3cd-/- female mice, including PD21 untreatment, PD21 after PMSG administration for 44 hours, PD21 after E2 administration for 72 hours, each group contained three biological repeats. The genes up/down-regulated significantly (not less than 2 fold change) in Pik3cd+/- mice after PMSG and E2 treatment were listed out as PMSG or E2 target genes, in which, those had not the same change trends in Pik3cd-/- were considered as Pik3cd-dependent genes.
2012-10-23 | E-GEOD-41746 | biostudies-arrayexpress
Project description:Transcriptional profiling of Sertoli cells of Monkey origin comparing infant Sertoli cells with pubertal Sertoli cells, isolated from Monkeys in which puberty was induced by extrinsic GnRH administration. Both Infant and pubertal cells were treated with FSH and Testosterone in vitro.
2015-06-01 | GSE57937 | GEO
Project description:Transcriptional profiling of Sertoli cells of Monkey origin comparing infant Sertoli cells with pubertal Sertoli cells, isolated from Monkeys in which puberty was induced by extrinsic GnRH administration. Both Infant and pubertal cells were treated with FSH and Testosterone in vitro.
2017-02-20 | GSE66129 | GEO
Project description:Imbalanced nutrition and exogenous gonadotrophins affect uterine function and morphology. In sheep, FSH-induced superovulation causes morphological alterations in the uterus, leading, i.e., to an increase in lipid droplets. Similarly, depending on diet, FSH treatment alters the expression of estrogen and progesterone receptors and of adhesion molecules. To broad the knowledge regarding imbalanced nutrition and carry-over effects of superovulation induced with FSH in the caruncular endometrium, deep RNA sequencing (NGS, RNA-Seq) was applied, followed by functional analysis. Ewes (n = 3-5/group) were divided as follows: control fed (CF), overfed (OF), and underfed (UF), and each group was further treated with FSH (superovulated (SOV)) or saline (negative controls (CONT)). Samples were collected on day 10 of diestrus. Furthermore, samples from CF_CONT were collected on day 5 to assess time-dependent changes. The 1484 differentially expressed genes (DEGs, P < 0.01, FDR < 0.05) identified between CF_CONT animals at days 5 and 10 were associated with increased immune activity and cellular metabolic processes and decreased cellular proliferation. In CONT animals, unbalanced nutrition (i.e., both OF and UF) was associated with enrichment of terms associated with cell adhesion and differentiation, immune response and angiogenesis. As for carry-over effects of FSH, higher number of DEGs were found for CF animals (1374), than in OF (168) or UF (18) samples, associated with dysregulation of cell cycle and hormonal sensitivity. The absence of genes concomitantly affected by SOV at all different feeding regimes suggested multidirectional and body condition-dependent effects of FSH in caruncular transcriptome. Both body condition and SOV appear to affect the homeostasis of the ovine caruncles, possibly involving the modulation of uterine receptivity.
2024-04-19 | GSE229048 | GEO
Project description:To unravel the genes that underpin the effects of FSH on spermatogenesis, testicular gene expression was compared in juvenile rats after FSH suppression to littermate controls. Based on existing data, it is apparent that the dominant function of FSH shifts between 9 and 18 day postpartum (dpp) during the first wave of spermatogenesis from driving Sertoli cell proliferation to support of germ cells. To enable comprehensive analysis of the impact of acute in vivo FSH suppression on Sertoli and germ cell development and to determine the genes that underpin these affects, FSH was selectively suppressed in Sprague Dawley rats by passive immunisation for 4 days prior to testis collection at 18 days postpartum. FSH suppression did not affect Sertoli cell proliferation at 18 dpp, but germ cell numbers were decreased at 18 dpp following FSH suppression, with a corresponding increase in germ cell apoptosis measured. Sixty transcripts were measured as changed at 18 dpp in response to 4 days of FSH suppression, as assessed using Affymetrix⢠microarrays. Some of these are known as Sertoli cell-derived FSH-responsive genes (e.g. StAR, cathepsin L, insulin-like growth factor binding protein-3), while others encode proteins involved in cell cycle and survival regulation (e.g. cyclin D1, scavenger receptor class B 1). These data demonstrate that FSH affects germ cells in vivo, by supporting germ cell viability at day 18. This model enabled identification of candidate genes that contribute to the FSH-mediated pathway by which Sertoli cells support germ cells. Experiment Overall Design: Sprague Dawley male rats at day 14 postpartum received either a rat FSH antibody raised in sheep or a control sheep immunoglobulin for 4 days. RNA was extracted from the testis and hybridisation on Affymetrix microarrays was conducted.
2008-06-11 | E-GEOD-10293 | biostudies-arrayexpress