Project description:Chondrocytes can potentially perceive mechanical stimuli via Piezo channels. We investigated the effect of the Piezo1 agonist Yoda1 on chondrocyte-like ATDC5 cells. Chondrocytes can potentially perceive mechanical stimuli via Piezo channels. We investigated the effect of the Piezo1 agonist Yoda1 on chondrocyte-like ATDC5 cells. Chondrocytes can potentially perceive mechanical stimuli via Piezo channels. We investigated the effect of the Piezo1 agonist Yoda1 on chondrocyte-like ATDC5 cells. Chondrocytes can potentially perceive mechanical stimuli via Piezo channels. We investigated the effect of the Piezo1 agonist Yoda1 on chondrocyte-like ATDC5 cells. We used microarray analysis to detail the global gene expression of ATDC5 cells in response to 6 hours of treatment with 5µM Yoda1.
Project description:Limb development is a well-established model for understanding cell fate decisions, and the formation of skeletal elements is coordinated through a sequence of events that control chondrogenesis spatiotemporally. It has been established that epigenetic control participates in cartilage maturation. Our research has shown for the first time that the inhibition of DNA methylation in interdigital tissue with 5-azacytidine results in the formation of an ectopic digit. This discovery suggested that DNA methylation dynamics could regulate the fate of cells between chondrogenesis and cell death during autopod development. We sequenced WGBS libraries from in-vivo interdigital tissue treated with 5-azacytidine or DMSO to identify genome-wide methylation changes during the early stages of 5-azacytidine-induced chondrogenesis.
Project description:Numerous studies have established a critical role for BMP signaling in skeletal development. In the developing axial skeleton, sequential SHH and BMP signals are required for specification of a chondrogenic fate in somitic tissue. A similar paradigm is thought to operate in the limb, but the signals involved are unclear. To investigate the nature of these signals we examined BMP action in mesenchymal populations derived from the early murine limb bud (~ E10.5). These populations exhibited a graded response to BMPs, in which early limb mesenchymal (EL) cells (from the distal hind limb) displayed an anti-chondrogenic response, whereas BMPs promoted chondrogenesis in older cell populations. To better understand the molecular basis of disparate BMP action in these various populations, gene expression profiling with Affymetrix microarrays was employed to identify BMP-regulated genes. These analyses showed that BMPs induced a distinct gene expression pattern in the EL cultures versus later mesenchymal limb populations (IM and LT). Mouse embryos at gestational age E10.5 were collected and various portions of the limb were micro-dissected. These led to the generation of three populations of cells, early (EL) limb mesenchymal cells from the distal half of the hind limb, an intermediate (IM) population derived from the distal 1/3 of the fore limb, and a later (LT) population from the proximal 2/3 of the fore limb. Mesenchymal cells were isolated and cultured with and without BMP4 treatment. RNA was extracted from cultures at either Day 0,1 or 2, labelled and hybridized to Affymetrix 430 2.0 microarrays. For each time point, RNA was collected from two biological replicates for each treatment condition.
Project description:We performed genome-wide transcriptome profiling in stable Kmt2d-/- (bi-allelic deletion of the catalytic SET domain) and Kmt2d+/+ ATDC5 chondrocyte cell lines 7 days after induction of differentiation and in stable Kmt2d-/- and Kmt2d+/+ undifferentiated ATDC5 cells.
Project description:Chondrogenic differentiation was induced in ATDC5 cells in the absence (RNAi) or presence of EGR1 (Early Growth Response Protein 1); RNA samples were collected at t=0, 2, 4.5, 8,16, 24, 72 hrs.
Project description:Five SAGE libraries were generated from A. thaliana leaf tissue collected at time points ranging from 30 minutes to one week of low temperature treatment (4°C). Over 240,000 high quality SAGE tags, corresponding to 16,629 annotated genes, provided a comprehensive survey of changes in the transcriptome in response to low temperature, from perception of the stress to acquisition of freezing tolerance. Keywords: SAGE; time course; stress response; cold acclimation; freezing tolerance