The G Protein-Coupled Receptor ADGRG6 Maintains Mouse Growth Plate Homeostasis by Negatively Regulating IHH Signaling
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ABSTRACT:
PROVIDER: PRJNA1150119 | ENA |
REPOSITORIES: ENA
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| SRR30311107.fastq.gz | Fastqsanger.gz | |||
| SRR30311108.fastq.gz | Fastqsanger.gz |
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Project description:ADGRG6 is a cartilage-enriched G protein-coupled receptor (GPCR). Using molecular mouse genetics and spatial transcriptomics approaches, we demonstrated that Adgrg6 has a vital role in regulating chondrocyte differentiation and growth plate homeostasis by positively regulating the formation and/or maintenance of the PTHrP (+) cell population and negatively regulating the IHH signaling in postnatal growth plates.
2024-09-19 | GSE275252 | GEO
Project description:The cartilage growth plate is essential for maintaining skeletal growth; however, the mechanisms governing postnatal growth plate homeostasis are still poorly understood. Using approaches of molecular mouse genetics and spatial transcriptomics applied to formalin-fixed, paraffin-embedded tissues, we show that ADGRG6/GPR126, a cartilage-enriched adhesion G protein-coupled receptor (GPCR), is essential for maintaining slow-cycling resting zone cells, appropriate chondrocyte proliferation and differentiation, and growth plate homeostasis in mice. Constitutive ablation of Adgrg6 in osteochondral progenitor cells with Col2a1Cre leads to a shortened resting zone, formation of cell clusters within the proliferative zone, and an elongated hypertrophic growth plate, marked by limited expression of parathyroid hormone-related protein (PTHrP) but increased Indian Hedgehog (IHH) signaling throughout the growth plate. Attenuation of smoothened-dependent hedgehog signaling restored the Adgrg6 deficiency-induced expansion of hypertrophic chondrocytes, confirming that IHH signaling can promote chondrocyte hypertrophy in a PTHrP-independent manner. In contrast, postnatal ablation of Adgrg6 in mature chondrocytes with AcanCreERT2, induced after the formation of the resting zone, does not affect PTHrP expression but causes an overall reduction of growth plate thickness marked by increased cell death specifically in the resting zone cells and a general reduction of chondrocyte proliferation and differentiation. Spatial transcriptomics reveals that ADGRG6 is essential for maintaining chondrocyte homeostasis by regulating osteogenic and catabolic genes in all the zones of the postnatal growth plates, potentially through positive regulation of SOX9 expression. Our findings elucidate the essential role of a cartilage-enriched adhesion GPCR in regulating cell proliferation and hypertrophic differentiation by regulation of PTHrP/IHH signaling, maintenance of slow-cycle resting zone chondrocytes, and safeguarding chondrocyte homeostasis in postnatal mouse growth plates.
| S-EPMC11523133 | biostudies-literature
Project description:In order to characterize mRNA expression in the growth plate, we microdissected postnatal rat growth plates into their constituent zones and used microarray analysis to assess the abundences of individual transcripts. Expression patterns of PTHrP and Ihh-related genes were confirmed using real-time PCR. Using a gli1-lacZ mouse, Gli1 expression, presumably representing Ihh signaling, was visualized during pre- and postnatal development.
2010-12-01 | GSE23432 | GEO
Project description:In order to characterize mRNA expression in the growth plate, we microdissected postnatal rat growth plates into their constituent zones and used microarray analysis to assess the abundences of individual transcripts. Expression patterns of PTHrP and Ihh-related genes were confirmed using real-time PCR. Using a gli1-lacZ mouse, Gli1 expression, presumably representing Ihh signaling, was visualized during pre- and postnatal development. Microdissection was used to collect individual growth plate zones from proximal tibiae of 1-wk rats and gene expression was analyzed using microarray.
2010-12-01 | E-GEOD-23432 | biostudies-arrayexpress
Project description:We performed single cell transcriptomic profiling of induced human pluripotent stem cells (iPSCs)-derived type 2 alveolar epithelial cells (iAT2). iPSCs stably expressed CRISPRi (dCas9-KRAB) under the control of doxycyline. iAT2s were transduced with a lentivirus expressing gRNA targeting the transcriptional start site of ADGRG6. Cells were treated with or without doxycyline to intiate CRISPRi-knockdown. Prior to capture, cells were labelled with hashing antibodies (HTO). Cells were captured for 10x Genomics Single cell capture (V3 protocol), and GEX and HTO libraries were sequenced. HTODemux function was used to demultiplex the samples. Knockdown of ADGRG6 caused cells to cluster separately from wild-type cells.
2023-09-20 | GSE223078 | GEO
Project description:The goal of the microarray analysis is to determine the redundant and distinct roles of Dhh and Ihh in ovarian functions
2018-05-30 | GSE109842 | GEO
Project description:Transcriptomes comparisons of Dhh KO, Ihh KO and Dhh/Ihh DKO ovaries
| PRJNA432169 | ENA
Project description:The purpose of this experiment was to obtain samples for mRNA analysis in IHH cells infected with Zaire Ebola virus and mutants: Zaire Ebola virus: This wild-type Ebola virus - strain Mayinga - was isolated from a fatal human case in Zaire (now known as the Democratic Republic of Congo) in 1976 Zaire Ebola virus, VP35 R312A possesses a R312A mutation in the VP35 protein. Zaire Ebola virus, delta sGP. Lacks the ability to produce non-structural protein, the secreted glycoprotein (sGP). Zaire Ebola virus, delta mucin. Lacks the mucin-like domain (MLD), which contains both N-linked and O-linked glycosylation sites, for the glycoproteins. Overview of Experiment: Cells: Immortalized Human Hepatocytes (IHH); seed 60,000 cells per well in a 24-well plate. Infected with a multiplicity of infection (MOI) of 0.5. After infection, 3x wash with PBS and replace with 5% FCS DMEM without NaPyr or NEAA. Time matched mocks done in triplicate from same cell stock as rest of samples. Time Points = 0, 6, 12, 24, 48, and 72 hrs post infection in triplicate.
2015-06-18 | E-GEOD-69943 | biostudies-arrayexpress
Project description:The purpose of this experiment was to obtain samples for mRNA analysis in IHH cells infected with Zaire Ebola virus and mutants: Zaire Ebola virus: This wild-type Ebola virus - strain Mayinga - was isolated from a fatal human case in Zaire (now known as the Democratic Republic of Congo) in 1976 Zaire Ebola virus, VP35 R312A possesses a R312A mutation in the VP35 protein. Zaire Ebola virus, delta sGP. Lacks the ability to produce non-structural protein, the secreted glycoprotein (sGP). Zaire Ebola virus, delta mucin. Lacks the mucin-like domain (MLD), which contains both N-linked and O-linked glycosylation sites, for the glycoproteins. Overview of Experiment: Cells: Immortalized Human Hepatocytes (IHH); seed 60,000 cells per well in a 24-well plate. Infected with a multiplicity of infection (MOI) of 0.5. After infection, 3x wash with PBS and replace with 5% FCS DMEM without NaPyr or NEAA. Time matched mocks done in triplicate from same cell stock as rest of samples. Time Points = 0, 6, 12, 24, 48, and 72 hrs post infection in triplicate.
2015-06-18 | E-GEOD-69942 | biostudies-arrayexpress
Project description:Purpose: Degenerative changes of the intervertebral disc are a leading cause of back pain and disability worldwide. However, precise mechanisms driving the initiation and progression of pathology have remained elusive. We find that adhesion G-protein coupled receptor Adgrg6 plays a critical role in maintaining postnatal intervertebral disc homeostasis. The goal of this study is to uncover early molecular changes in Adgrg6-defeicnet intervertebral discs prior to overt histopathology. Methods: Intervertebral disc mRNA profiles of 20-day-old wild type and Col2Cre; Adgrg6f/f mutant mice were generated by deep sequencing in triplicates. Cutadapt and perl scripts were used to remove the reads that contained adaptor contamination, low quality bases and undetermined bases. Then sequence quality was verified using FastQC. We used HISAT2 to map reads to the genome of Mus Musculus (GRCm38.88). The mapped reads of each sample were assembled using StringTie. Then, all transcriptomes from biological samples were merged to reconstruct a comprehensive transcriptome using perl scripts and gffcompare. After the final transcriptome was generated, StringTie and Ballgown was used to estimate the expression levels of all transcripts. StringTie was used to perform expression level for mRNAs by calculating FPKM. The differentially expressed mRNAs were selected with log2 (fold change) >1 or log2 (fold change) <-1 and with statistical significance (p value < 0.05) by R package Ballgown. Results: We found 884 differential expressed genes with statistical significance (p value < 0.05, and with a more stringent cut-off adjusted p value <0.05 and fold-change >2, we observed 42 differential expressed genes. Enriched pathways and biological processes using gene ontology (GO) terms included extracellular matrix, positive regulation of fibroblast proliferation, extracellular matrix structural constituent conferring tensile strength, regulation of tyrosine phosphorylation of STAT protein, and ion transport. We also find significantly increased expression of fibrotic collagens, induced expression of some Suppressor of Cytokine Signaling (SOCS) genes, and dysregulated expression of some components associated with ion transport system. Several of the significantly upregulated genes are associated with biomarkers or risk of lumbar disc degeneration and osteoarthritis in humans or animal models. Conclusions: Our study provides detailed analysis of intervertebral disc transcriptomes from both wild type and Adgrg6-deficient mice, with three biological replicates, prior to overt histopathology. Our transcriptomic analysis demonstrated a robust dysregulation of several important pathways and components of the intervertebral disc homeostasis, including induction of fibrotic gene expression, alteration of ion transport component, as well as changes in some chondrogenic and catabolic factors, prior to the onset of histopathology and disc degeneration. These data strongly suggest that ADGRG6 signaling is a critical factor for the maintenance of healthy gene expression profiles in the IVD.
2019-03-17 | GSE128402 | GEO