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Transcriptional analysis of tenocytes from mice with a targeted deletion of scleraxis

ABSTRACT: We report RNA sequencing data from tenocytes cultured from the tail tendons of adult male mice in the C57Bl/6 background that either have the scleraxis gene (Scx+) or mice in which scleraxis has been deleted using CreERT2 driven from the Rosa26 locus using 4-hydroxytamoxifen (4HT).

ORGANISM(S): Mus musculus

PROVIDER: GSE145863 | GEO | 2020/02/24

REPOSITORIES: GEO

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Transcriptional analysis of tendons from mice with a targeted deletion of scleraxis

Project description:We report RNA sequencing data from the plantaris tendons of adult male mice in the C57Bl/6 background that either have the scleraxis gene (Scx+) or mice in which scleraxis has been deleted using CreERT2 driven from the Rosa26 locus (Scx-). Mice in which scleraxis was deleted were created by crossing scleraxis-floxed mice with Rosa26-CreERT2 mice. Rosa26-CreERT2 mice that did not have a floxed scleraxis allele served as controls. Mice were treated with tamoxifen for 5 days to induce recombination at the scleraxis locus, and then subjected to a synergist ablation procedure in which the Achilles tendon is removed, resulting in compensatory growth of the plantaris tendon. Mice were analyzed either 7 or 14 days after synergist ablation. Control mice that did not undergo tamoxifen treatment or synergist ablation were also studied (NOC).

2020-01-31 | GSE139493 | GEO

RNA-seq analysis of equine tenocytes exposed to siRNA targeting scleraxis (Scx) to identify novel targets of Scx-mediated mechanotransduction

Project description:Equine tenocytes were exposed to an siRNA targeting the transcription factor scleraxis (Scx) or a scramble control and maintained under cyclic strain. The resulting transcriptomes were then used for RNA-seq analysis. The goal of this study was to identify novel ways in which Scx facilitates mechanotransduction in tenocytes in order to better understand tenocyte biology.

2018-08-02 | GSE110567 | GEO

The transcription factor Scleraxis differentially regulates gene expression in tenocytes isolated at different developmental stages

Project description:Purpose:- RNA sequencing was performed on adult and fetal equine tenocytes expressing a specific short hairpin RNA against scleraxis (shSCX) versus a non-target (NT) scrambled control Methods:- mRNA profiles of four independent biological lines of adult and fetal NT and shSCX expressing tenocytes were generated by RNA sequencing, using Illumina NovaSeq6000. The sequence reads that passed quality filters were analyzed using the pseudoaligner Salmon and differential expression quantified using DESeq2. Gene ontology analysis was then conducted using Panther. Pathway analysis was conducted using GeneAnalytics. Network analysis was conducted using the STRING plugin in the Cytoscape app. Results:- SCX knockdown had a larger effect on gene expression in fetal tenocytes, effecting 477 genes in comparison to the 183 genes effected in adult tenocytes, indicating that SCX-dependent processes may differ in these two developmental stages.

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Scleraxis-lineage cells are required for tendon homeostasis and their depletion induces an accelerated extracellular matrix aging phenotype

Project description:Aged tendons have disrupted homeostasis, increased injury risk, and impaired healing capacity. Understanding mechanisms of homeostatic disruption is crucial for developing therapeutics to retain tendon health through the lifespan. Here, we developed a novel model of accelerated tendon extracellular matrix (ECM) aging via depletion of Scleraxis-lineage (ScxLin) cells in young mice (DTR). DTR recapitulates many aspects of tendon aging including comparable declines in cellularity, alterations in ECM structure, organization, and composition. Single cell RNA-sequencing demonstrated a conserved decline in tenocytes associated with ECM biosynthesis in aged and DTR tendons, identifying the requirement for ScxLin cells during homeostasis. However, the remaining cells in aged and DTR tendons demonstrate functional divergence. Aged tenocytes become pro-inflammatory and lose proteostasis. In contrast, DTR tenocytes demonstrate enhanced remodeling capacity. Collectively, this study defines DTR a novel model of accelerated tendon ECM aging and identifies novel biological intervention points to maintain tendon function through the lifespan.

2023-02-03 | PXD037230 | Pride

Transcriptional analysis of tendons from mice with a tenocyte-specific deletion of IGF1R

Project description:We report RNA sequencing data from the plantaris tendons of adult male mice in the C57Bl/6 background that either have the IGF1 receptor (IGF1R) present in their tendons (Scx:IGF1R+) or mice in which IGF1R has been deleted in tenocytes expressing scleraxis (Scx:IGF1R-). Mice were created by crossing ScxCreERT2 mice with IGF1R flox/flox mice. Mice were treated with tamoxifen for 5 days to induce recombination at the IGF1R allele, and then subjected to a synergist ablation procedure in which the Achilles tendon is removed, resulting in compensatory growth of the plantaris tendon. Mice were analyzed either 7 or 14 days after synergist ablation. Control mice that did not undergo tamoxifen treatment or synergist ablation were also studied.

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Transcriptional analysis of tendons and tenocytes from mice with a targeted deletion of scleraxis

Project description:Transcriptional analysis of tendons and tenocytes from mice with a targeted deletion of scleraxis

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RNA-seq analysis of equine tenocytes exposed to siRNA targeting scleraxis (Scx) to identify novel targets of Scx-mediated mechanotransduction

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Proteomic analysis of scleraxis mutant tendon

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2021-09-08 | PXD017944 | Pride

Tenogenic induction from induced pluripotent stem cells unveils the trajectory towards tenocyte differentiation.

Project description:The musculoskeletal system is integrated by tendons that are characterized by expression of a functionally important transcription factor Scleraxis (Scx). To date, there has been no solid culture system for tenogenic differentiation. Here we developed the tenocyte induction method using induced pluripotent stem cells established from ScxGFP transgenic mice by monitoring fluorescence that reflects a series of dynamic differentiation process. Among several developmentally relevant factors, TGF-2 was the most potent inducer for differentiation of tenocytes expressing Tenomodulin, a mature differentiation marker. Single-cell RNA sequencing (scRNA-seq) revealed 11 distinct clusters including mature tenocyte population and tenogenic differentiation trajectory, which well recapitulates in vivo developmental process. Dataset of scRNA-seq and transcriptome of Scx deficient tendon enabled identification of Scx-related genes in which tendon-related genes are enriched. Collectively, this study leads to a new opportunity for tendon research and provides further insight into mechanistic understanding of the differentiation pathway to a tenogenic fate.

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Project description:We report RNA sequencing data from tenocytes treated with IGF1. Tenocytes were obtained from the tail tendons of adult C57Bl/6 mice via collagenase digestion. Tenocytes were grown to 60% confluence, and then treated with 100ng/mL of recombinant IGF1 for a period of 0, 1, 2, 6, or 24 hours. Experiments were conducted in quadruplicate. RNA was isolated and prepared for RNA sequencing.

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