Overexpression of Hoxd4 transcription factor alters transcriptional profiles in mouse chondrocytes at E18.5
Ontology highlight
ABSTRACT:
ORGANISM(S): Mus musculus
SUBMITTER: Claudia Kruger
PROVIDER: E-GEOD-18991 | biostudies-arrayexpress |
REPOSITORIES: biostudies-arrayexpress
Similar Datasets
Project description:Homeobox genes of the Hox class are required for proper patterning of skeletal elements and play a role in cartilage differentiation. In transgenic mice with overexpression of Hoxc8 during cartilage development, we observed severe defects, namely physical instability of cartilage, accumulation of immature chondrocytes, and decreased maturation to hypertrophy. To define the molecular basis underlying these defects, we performed gene expression profiling using the Affymetrix microarray platform. Embryos were dissected from four different mouse litters at E18.5. Embryos from each litter were grouped according their genotype: TA=control; TR=transgenic.
2010-10-15 | E-GEOD-18992 | biostudies-arrayexpress
Project description:This SuperSeries is composed of the following subset Series: GSE18991: Overexpression of Hoxd4 transcription factor alters transcriptional profiles in mouse chondrocytes at E18.5 GSE18992: Overexpression of Hoxc8 transcription factor alters transcriptional profiles in mouse chondrocytes at E18.5!Series_overall_design = Refer to individual Series Refer to individual Series
2010-10-15 | E-GEOD-19002 | biostudies-arrayexpress
Project description:Endochondral bone formation is orchestrated by growth factors produced by chondrocytes and deposited in cartilage matrix. Some of these factors were identified, but their complete list and relationship remains unknown. In this work the growth factors were isolated from calcified cartilage of costochondral junctions. The isolation involved hyaluronidase digestion, guanidinium hydrochloride (GuHCl) extraction, HCl decalcification and GuHCl extraction of decalcified matrix. Growth factors were purified by heparin chromatography and estimated by enzyme-linked immunosorbent assay (ELISA) or used for protein sequence analysis. Bone morphogenetic protein-7 (BMP-7), Growth/differentiation factor 5 (GDF-5), and NEL-like protein 1 (NELL-1) quantitatively dominated in material obtained in all steps of isolation. During ossification perivascular cells and septoclasts enter the cartilage and release growth factors stimulating osteoclastogenesis. Osteoclasts dissolve calcified cartilage and transport released factors needed for stimulation of osteoprogenitor cells. Presence of BMP-7, GDF-5 and NELL-1 at the site of initial bone formation may suggest that their synergistic action favors osteogenesis.
2021-05-03 | PXD021781 | Pride
Project description:A comprehensive analysis of Sox9 binding profiles in developing chondrocytes identified marked enrichment of an AP-1-like motif (Ohba et al. 2015). Here, we have explored the functional interplay between Sox9 and AP-1 in mammalian chondrocyte development. Among AP-1 family members, Jun and Fosl2 were highly expressed within prehypertrophic and early hypertrophic chondrocytes. Chromatin immunoprecipitation followed by DNA sequencing (ChIP-seq) showed a striking overlap in Jun- and Sox9-bound regions throughout the chondrocyte genome, a reflection of direct binding of each factor to target motifs in shared enhancers, and physical interactions of AP-1 with Sox9. In vitro expression analysis indicates that direct co-binding of Sox9 and AP-1 at target motifs enhanced target gene expression, while protein-protein interactions suppressed AP-1- and Sox9-driven transcription. Analysis of prehypertrophic chondrocyte removal of Sox9 demonstrated Sox9 was essential for hypertrophic chondrocyte development, while in vitro and ex vivo analyses showed AP-1 promotes chondrocyte hypertrophy. Sox9 and Jun co-bound and co-activated a Col10a1 enhancer in Sox9 and AP-1 motif-dependent manners consistent with their combined action promoting hypertrophic gene expression. Together, the data support a model where AP-1-family members promote Sox9-action in the transition of chondrocytes to a terminal hypertrophic program. Intersection of ChIP-seq data from Sox9 and AP-1 factor Jun, RNA-seq data from developing rib chondrocytes and Col10a1mCherry positive hypertrophic chondrocytes in neonatal mice to uncover regulation of Sox9 by AP-1 factors during chondrocyte hypertrophy.
2016-02-29 | E-GEOD-73372 | biostudies-arrayexpress
Project description:BBF2H7 (BBF2 human homolog on chromosome 7), an ER-resident basic leucine zipper transcription factor, is activated in response to ER stress and abundantly expresses in chondrocytes. While BBF2H7 is widely expressed in many tissues and organs, the most intense signals were detected in the proliferating zone of the cartilage. We compared gene expressions in primary cultured chondrocytes prepared from rib cartilage between WT and BBF2H7-/- mice at E18.5. Primary cultured chondrocytes were prepared from E18.5 rib cartilage of WT and BBF2H7-/- mice. Chondrocytes were isolated using 0.2% collagenase D (Roche) after adherent connective tissue was removed by 0.2% trypsin (Sigma) and collagenase pretreatment. Isolated chondrocytes were maintained in α-MEM (Gibco) supplemented with 10% FCS and 50 µg/mL ascorbic acid. Adenovirus vectors expressing the mouse p60 BBF2H7 (1-377 aa, BBF-N) were constructed with the AdenoX Expression system (Clontech), according to the manufacturerâs protocol. The cells were infected with adenoviruses 30 h before analysis. We compared gene expressions in primary cultured chondrocytes prepared from rib cartilage between WT and BBF2H7-/- mice at E18.5 using a microarray and various genes associated with protein secretory pathway and ER biogenesis were significantly down-regulated in BBF2H7-/- chondrocytes. We infected primary cultured chondrocytes prepared from BBF2H7-/- mice with adenovirus expressing p60 BBF2H7. Several genes were up-regulated and we picked up them as the direct target of BBF2H7.
2010-05-05 | E-GEOD-18052 | biostudies-arrayexpress
Project description:Objectives: To identify similarities and differences in gene expression data in the MEK/ERK and PI3K pathways and to determine how histone modification affects these same pathways. Goal: To identify and functionally characterize novel targets of these signaling pathways in the context of chondrocyte differentiation. Experiment Overall Design: Cartilage derived from the hind limbs of CD1 mice staged at 15.5 days of embryonic development were dissected and plated in primary chondrocyte monolayer cultures. Cells were treated with MEK/ERK pathway inhibitors (SB202190 and UO126), a PI3K inhibitor (LY294002), a histone deacetylase inhibitor (Trichostatin A) and the vehicle control (DMSO) for 24 hours. Total RNA was isolated from these samples and hybridized to Affymetrix MOE 430 2.0 chips at the London Regional Genomics facility. Experiment Overall Design: A total of 15 genechips were analyzed between 4 treatments and the vehicle (DMSO) control. Three biological replicates per treatment were executed. Experiment Overall Design: Experiment Overall Design:
2009-05-11 | E-GEOD-15069 | biostudies-arrayexpress
Project description:MicroRNAs expression profiling of human nucleus pulposus cells derived from patients with disc degeneration in comparison with those derived from patients with scoliosis as control. Two-condition experiment: control nucleus pulposus cells vs. degenerative nucleus pulposus cells. Biological replicates: 3 control, 3 degenerated, independently harvested. Four replicates per array. The supplementary file 'GSE19943_fold_pvalue.txt' contains fold-changes and p-values.
2012-01-01 | E-GEOD-19943 | biostudies-arrayexpress
Project description:Sixteen samples; 6 nucleus pulposus, 6 annulus fibrosus and 4 articular cartilage were compared to find transcriptional differences.
2010-12-27 | E-MEXP-2936 | biostudies-arrayexpress
Project description:Background: Glucocorticoids (GCs) are widely used anti-inflammatory drugs. While useful in clinical practice, patients taking GCs often suffer from skeletal side effects including growth retardation and decreased bone quality in adults. On a physiological level, GCs have been implicated in the regulation of chondrogenesis and osteoblast differentiation, as well as maintaining homeostasis in cartilage and bone. We identified the glucocorticoid receptor (GR) as a potential regulator of chondrocyte hypertrophy in a microarray screen of primary limb bud mesenchyme micromass cultures. Some targets of GC regulation in chondrogenesis are known, but the global effects of pharmacological GC doses on chondrocyte gene expression have not been comprehensively evaluated. Results: This study systematically identifies a spectrum of GC target genes in embryonic growth plate chondrocytes treated with synthetic GR agonist, dexamethasone (DEX), at 6 and 24 hrs. Conventional analysis of this data set and gene set enrichment analysis (GSEA) was performed. Transcripts associated with metabolism were enriched in the DEX condition along with extracellular matrix genes. In contrast, a subset of growth factors and cytokines were negatively correlated with DEX treatment. Comparing DEX-induced gene expression data to developmental changes in gene expression in micromass cultures revealed an additional layer of complexity in which DEX maintains the expression of certain chondrocyte marker genes while inhibiting factors that promote vascularization and ultimately ossification of the cartilaginous template. Conclusions: Together, these results provide insight into the mechanisms and major molecular classes functioning downstream of DEX in primary chondrocytes monolayers. In addition, comparison of our data with microarray studies of DEX treatment in other cell types demonstrated that the majority of DEX effects are tissue-specific. This study provides novel insights into the effects of pharmacological GC on chondrocyte gene transcription and establishes the basis for subsequent functional studies. Experiment Overall Design: Primary chondrocytes isolated from the tibiae, humeri and femurs of 15.5 day old mice are treated with 10EXP-7M DEX or an equal volume of the DMSO vehicle for 24 hrs. Total RNA is isolated after 6 hrs and 24 hrs of treatment. Samples from three independent experiments independent time courses were hybridized to Affymetrix MOE 430 2.0 mouse chips. Experiment Overall Design: Number of time points: 2 (6hr and 24 hr) Experiment Overall Design: Number of treatments: 2 (DEX and the vehicle control) Experiment Overall Design: Number of Samples: 3 replicates per time point per treatment Experiment Overall Design: Affymetrix chip: MOE 430 2.0 Experiment Overall Design: Cell type: Primary chondrocyte Experiment Overall Design: Tissue or origin: Tibiae, humerus, femur Experiment Overall Design: Species E15.5 mice Experiment Overall Design: Samples: Total RNA
2008-06-15 | E-GEOD-7683 | biostudies-arrayexpress
Project description:Prospectively isolated and characerized skeletal progenitor lineages 3 repeats per FACS sorted population
2015-01-29 | E-GEOD-64447 | biostudies-arrayexpress