Glucocorticoid transcriptome in the hippocampus of wild-type and GRA knockin mice
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ABSTRACT:
ORGANISM(S): Mus musculus
PROVIDER: GSE298763 | GEO | 2026/06/08
REPOSITORIES: GEO
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Project description:The glucocorticoid receptor (GR) regulates gene expression upon activation by glucocorticoid (GC) hormones. In zebrafish, two GR splice variants exist: the canonical GR α-isoform (GRα), and the GR β-isoform (GRβ). The exact function of GRb remains elusive. We have investigated the transcriptional role of GRa and GRb in the zebrafish embryo model by injecting mebryos with two splice-blocking morpholinos (one leading to knockdown of both GR α- and β-isoforms, and another targeting the alternative splicing of the GR pre-mRNA in favor of the GR β-isoform) and with GRβ mRNA (resulting in specific GRβ overexpression). Transcriptome profiling was performed on total RNA isolated from 30 hpf embryos. Our results show that GRb does not act as a dominant-negative inhibitor of GRa, and that GRa regulates two distinct gene clusters, which are mainly involved in the metabolism of the embryo.
2014-11-18 | GSE63360 | GEO
Project description:The glucocorticoid receptor (GR) regulates gene expression upon activation by glucocorticoid (GC) hormones. In zebrafish, two GR splice variants exist: the canonical GR M-NM-1-isoform (GRM-NM-1), and the GR M-NM-2-isoform (GRM-NM-2). The exact function of GRb remains elusive. We have investigated the transcriptional role of GRa and GRb in the zebrafish embryo model by injecting mebryos with two splice-blocking morpholinos (one leading to knockdown of both GR M-NM-1- and M-NM-2-isoforms, and another targeting the alternative splicing of the GR pre-mRNA in favor of the GR M-NM-2-isoform) and with GRM-NM-2 mRNA (resulting in specific GRM-NM-2 overexpression). Transcriptome profiling was performed on total RNA isolated from 30 hpf embryos. Our results show that GRb does not act as a dominant-negative inhibitor of GRa, and that GRa regulates two distinct gene clusters, which are mainly involved in the metabolism of the embryo. This microarray study was designed to determine the effect of knockdown and overexpression of GRa and GRb. For this purpose, wild type (AB/TL) zebrafish embryos were injected at the 1-2 cell stage with a standard control morpholino (SC-MO), a splice-blocking morpholino leading to knockdown of both GR M-NM-1- and M-NM-2-isoforms (MO1), a splice-blocking morpholino targeting the alternative splicing of the GR pre-mRNA in favor of the GR M-NM-2-isoform (MO2), or GRb mRNA. At 24 hpf, each group was treated with dexamathasone (or vehicle) for 6 hr. This resulted in 8 treament groups: SC-MO/veh, SC-MO/dex, MO1/veh, MO1/dex, MO2/veh, MO2/dex, GRb mRNA/veh, GRb mRNA/dex. After the incubation period, embryos were collected and RNA was isolated from 20 embryos per treatment group (30 hpf). Three individual experiments were performed, and the resulting triplicate samples were analyzed by microarray, using a common reference approach.
2014-11-18 | E-GEOD-63360 | biostudies-arrayexpress
Project description:Glucocorticoids are primary stress hormones and their synthetic derivatives are widely used clinically. The therapeutic efficacy of these steroids is limited by severe side effects and glucocorticoid resistance. Multiple glucocorticoid receptor (GR) isoforms are produced by alternative translation initiation; however, the role individual isoforms play in controlling tissue-specific responses to glucocorticoids in vivo is unknown. We have generated knockin mice that exclusively express the most active receptor isoform, GR-C3. The GR-C3 knockin mice die at birth due to respiratory distress but can be completely rescued by antenatal glucocorticoid administration. To evaluate the GR-C3 transcriptome, we prepared mouse embryonic fibroblasts (MEFs) from E12.5 wild-type (WT) and GR-C3 knockin embryos and treated the cells with vehicle or the synthetic glucocorticoid Dexamethasone (Dex) for 6 hours. The GR-C3 isoform was found to have a markedly different gene-regulatory profile than GR in WT MEFs.
2018-04-23 | GSE103675 | GEO
Project description:Glucocorticoids are primary stress hormones and their synthetic derivatives are widely used clinically. The therapeutic efficacy of these steroids is limited by severe side effects and glucocorticoid resistance. Multiple glucocorticoid receptor (GR) isoforms are produced by alternative translation initiation; however, the role individual isoforms play in controlling tissue-specific responses to glucocorticoids in vivo is unknown. We have generated knockin mice that exclusively express the most active receptor isoform, GR-C3. The GR-C3 knockin mice die at birth due to respiratory distress but can be completely rescued by antenatal glucocorticoid administration. Rescued GR-C3 mice exhibited alterations in circadian rhythm in a sexually dimorphic manner and in sensitivity to lipopolysaccharide (LPS)-induced endotoxemia. To evaluate the ability of glucocorticoids to protect against LPS-induced inflammation, we measured gene expression in spleens from WT and rescued GR-C3 knockin mice that had been treated with vehicle or LPS for 3 and 24 hours. The GR-C3 isoform was found to be deficient in its ability to repress a large cohort of immune and inflammatory genes.
2018-05-08 | GSE103721 | GEO
Project description:Glucocorticoids regulate diverse physiologic processes and synthetic derivatives of these natural hormones are widely used in the treatment of inflammatory diseases. However, chronic administration often triggers insensitivity and serious side effects including osteoporosis. The underlying mechanisms regulating these side effects are not completely understood. We report here that human osteosarcoma U-2 OS bone cells lacking the glucocorticoid receptor (GR) are resistant to glucocorticoid killing whereas the expression of wild-type GR activates an apoptotic program. Furthermore, we show that the translationally generated GR isoforms from a single GR gene have distinct abilities to induce apoptosis in these cells. Only cells expressing selective GR isoforms underwent apoptosis upon dexamethasone treatment and microarray analysis demonstrated that GR isoforms selectively stimulated the expression of pro-apoptotic enzymes such as caspase 6 and granzyme A. Chromatin immunoprecipitation assays further revealed that GR isoform-dependent induction of pro-apoptotic genes is likely due to selective coregulator recruitment and chromatin modification. Together, these findings provide evidence for a direct apoptotic effect of glucocorticoids on bone cells via selective GR isoforms and delineate multiple molecular components involved in tissue-specific glucocorticoid-induced bone cell apoptosis. Keywords: time course, isoform comparison
2007-12-20 | GSE6711 | GEO
Project description:The glucocorticoid receptor (GR/NR3C1), a ligand-activated transcription factor belonging to the nuclear receptor (NR) superfamily, is vital for life. Like other NRs, GR features an intrinsically disordered N-terminal domain (NTD) followed by a highly conserved DNA-binding domain (DBD) and a hormone- or ligand-binding domain (LBD), separated by a long linker. While the receptor's quaternary structure is critical for its transcriptional activity, its precise multimeric organization has remained elusive. Here, we conducted a comprehensive crosslinking-mass spectrometry (XL-MS) analysis using cleavable crosslinkers (DSSO and DSBU) to investigate the quaternary structure of the GR DBD-linker-LBD tandem. Our results strongly support a model of non-canonical receptor multimerization, revealing that GR monomers arrange in a parallel fashion through interactions involving residues within the LBD's L1-3 loop (evidenced by XLs K576-K576’ and K579-K579’). These parallel, non-canonical dimers then further associate to form dimers-of-dimers, mediated by contacts within the H9/L9-10/H10 region (e.g., K695-K695’). This study provides unambiguous evidence for a novel GR multimerization mechanism and has important implications for the design of glucocorticoids with fewer side effects.
2025-10-27 | PXD065782 | Pride
Project description:The glucocorticoid (GC) receptor (GR) is a ligand-induced transcription factor regulating metabolism and inflammation. While homologous downregulation of ligand-bound GR limits the response, it also reduces hormone responsiveness resulting in glucocorticoid resistance. Regulatory mechanisms that refine this equilibrium are less understood. Here, we have identified seven lysine acetylation sites in the amino terminal domain of GR, lysine 154 within the regulatory AF-1 region being the dominant acetyl-acceptor. GR-Lys154 acetylation is mediated by p300/CBP in an exclusively agonist-induced manner and correlates with nuclear translocation and transcriptional activity. GR-Lys154 acetylation is removed by the NAD+ dependent deacetylase, SIRT1, indicating dynamic regulation of this mark. Notably, agonist-binding to both the wild type and acetylation-deficient mutant GR elicits similar short-term target gene expression. In contrast, upon extended treatment, the acetylation-deficient mutant shows impaired ubiquitination and higher protein stability with a concomitant increase in chromatin association and transactivation. Taken together, p300/CBP and SIRT1 fine-tune glucocorticoid response by siphoning acetylated GR for proteasomal degradation while GR deacetylation sustains the transcriptional cascade for prolonged periods.
2024-01-16 | GSE236739 | GEO
Project description:Post-traumatic stress disorder (PTSD) is a maladaptive and debilitating psychiatric disorder with limited intervention targets. FRMD5, a scaffold protein, serves as molecular hub involved in diverse biological processes. This study elucidates a tissue-specific mechanism via shorter isoform of FRMD5, characterized by condensate formation and special interactive features, on modulation of glucocorticoid receptor (GR) transcriptional functions. Using a comprehensive set of techniques, we observed a reduction in stress-induced behaviors, highlighting the crucial role of shorter isoform of FRMD5. Further investigations revealed that shorter isoform of FRMD5 forms condensates in nuclear and interacts with a set of proteins involved in GR protein complex and transcriptional cofactors. Besides, FRMD5 accelerates the nuclear translocation and modulates transcriptional functions of GR. Systemic administration of peptides that disrupt FRMD5 functions led to transcriptional functions changes of GR and decreased stress-induced behaviors in mice. Overall, this study uncovers an intrinsic mechanism modulating transcriptional functions of GR via specific FRMD5 isoform, identifying a potential intervention target for PTSD.
2026-01-15 | GSE314512 | GEO
Project description:Glucocorticoid Regulated Transcriptome in Wild-type and GR-C3 Knockin Mouse Embryonic Fibroblasts
| PRJNA404034 | ENA
Project description:Glucocorticoids regulate diverse physiologic processes and synthetic derivatives of these natural hormones are widely used in the treatment of inflammatory diseases. However, chronic administration often triggers insensitivity and serious side effects including osteoporosis. The underlying mechanisms regulating these side effects are not completely understood. We report here that human osteosarcoma U-2 OS bone cells lacking the glucocorticoid receptor (GR) are resistant to glucocorticoid killing whereas the expression of wild-type GR activates an apoptotic program. Furthermore, we show that the translationally generated GR isoforms from a single GR gene have distinct abilities to induce apoptosis in these cells. Only cells expressing selective GR isoforms underwent apoptosis upon dexamethasone treatment and microarray analysis demonstrated that GR isoforms selectively stimulated the expression of pro-apoptotic enzymes such as caspase 6 and granzyme A. Chromatin immunoprecipitation assays further revealed that GR isoform-dependent induction of pro-apoptotic genes is likely due to selective coregulator recruitment and chromatin modification. Together, these findings provide evidence for a direct apoptotic effect of glucocorticoids on bone cells via selective GR isoforms and delineate multiple molecular components involved in tissue-specific glucocorticoid-induced bone cell apoptosis. Experiment Overall Design: U-2 OS cells were transfected with the BD Clontech pTET-OFF regulatory plasmid to establish the U-OFF parental cell line. MluI and EcoRV ends were generated onto the coding region of hGR-alpha, -A, -B, -C, or -D using PCR amplification of the pCMVhGR-alpha or -A plasmid. The pTRE2hyg vector was digested with MluI and EcoRV and the two DNAs were ligated to form the pTRE2hGR-alpha,-A, -B, -C, or -D plasmid. Each DNA construct was individually transfected into the U-OFF cells and clones were selected which stably expressed either hGR-alpha, -A, -B, -C, or -D using 200 microg/ml of geneticin and 500 mircorg/ml of hygromycin. Several clones were obtained for each receptor, and the receptor levels were compared using western blot analyses. In these cell lines, the expression of hGR can be repressed by the addition of tetracycline or the derivative doxycycline to the media. U-2 OS (human osteosarcoma) cells were maintained in DMEM/F-12 supplemented with 10% FCS:CS, 2 mM glutamine and pen-strep and selected clones were maintained in the same media with the addition of 200 microg/ml Geneticin and 200 mircrog/ml hygromycin. All cells were maintained in a humidified, 5% CO2 atmosphere. Cells were either not treated (0 hours) or were treated with 100 nM dexamethasone for 6, 12, or 24 hours. Experiment Overall Design: Total RNA was amplified using the Agilent Low RNA Input Fluorescent Linear Amplification Kit protocol. Starting with 500ng of total RNA, Cy3 labeled cRNA was produced according to manufacturer's protocol. For each sample, 1.5ug of Cy3 labeled cRNAs were fragmented using the Agilent In Situ Hybridization Kit protocol. Hybridizations were performed for 17 hours in a rotating hybridization oven at 65 degrees at 4 RPM. Slides were washed with 6X SSPE + 0.005% N-lauroyl sarcosine for 1 minute then 0.06X SSPE + 0.005% N-lauroyl sracosine for 1 minute at 37 degrees. The slides were dried by slowly removing from second wash solution and then scanned with an Agilent G2565 Scanner (10micron and with XDR) and processed with Agilent Feature Extraction v9.1. The resulting files were imported into the Rosetta Resolver system (Version 6.0). This system performs data pre-processing and error modeling.
2008-06-15 | E-GEOD-6711 | biostudies-arrayexpress