ABSTRACT: Comparison of human prepuberal articular and growth plate cartilage Total RNA was isolated from healthy human prepuberal tibiae and femurs donors
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.
Project description:To explore the mechanisms responsible for spatial and temporal regulation of the growth plate, we microdissected postnatal rat growth plates into their constituent zones and then used microarray analysis to characterize the changes in gene expression that occur as chondrocytes undergo spatially-associated differentiation and temporally-associated senescence. Microdissection was used to collect individual growth plate zones from proximal tibiae of 1-wk rats and the proliferative and early hypertrophic zones of growth plates from 3-, 6-, 9-, and 12-wk rats and gene expression was analyzed using microarray.
Project description:We used laser capture microdissection to isolate different zones of the articular cartilage from proximal tibiae of 1-week old mice, and used microarray to analyze global gene expression. Bioinformatic analysis corroborated previously known signaling pathways, such as Wnt and Bmp signaling, and implicated novel pathways, such as ephrin and integrin signaling, for spatially associated articular chondrocyte differentiation and proliferation. In addition, comparison of the spatial regulation of articular and growth plate cartilage revealed unexpected similarities between the superficial zone of the articular cartilage and the hypertrophic zone of the growth plate.
Project description:Gene expression profiles of bipolar disorder (BD) patients were assessed during both a manic and a euthymic phase and compared both intra-individually, and with the gene expression profiles of controls. 11 BD patients were assessed in their manic as well as in their euthymic phase. Comparison of gene expression BD euthymic vs. controls, BD manic vs. controls, and intra-patient comparison BD euthymic vs. BD manic.
Project description:Purpose: Next-generation sequencing (NGS) has revolutionized systems-based analysis of cellular pathways. The goals of this study are to evaluate POT1a deleted MSPCs functions by using NGS-derived MSPCs transcriptome profiling (RNA-seq) in vitro and vivo. Methods: In vitro, bone marrow cells flushed out from mouse femurs and tibiae were cultured, and adherent cells were collected. CD51+/PDGFRɑ+ cells were sorted and further cultured, and Cre Recombinase Gesicles were applied. After 48 hours culture, tdTomato fluorescently colored cells were re-sorted, cultured for about 4 weeks, and used for gene analyses. In vivo, bone marrow cells weer flushed out from mouse femurs and tibiae, and the flow-through fraction with tdTomato signals was sorted as an MSPC-enriched population. Results: Gene set enrichment analysis of the differentially expressed genes revealed that several pathways related to bone development were significantly affected. Conclusions: POT1a is essential for bone marrow MSPCs to maintain osteo- differentiation potential.
Project description:We used laser capture microdissection to isolate different zones of the articular cartilage from proximal tibiae of 1-week old mice, and used microarray to analyze global gene expression. Bioinformatic analysis corroborated previously known signaling pathways, such as Wnt and Bmp signaling, and implicated novel pathways, such as ephrin and integrin signaling, for spatially associated articular chondrocyte differentiation and proliferation. In addition, comparison of the spatial regulation of articular and growth plate cartilage revealed unexpected similarities between the superficial zone of the articular cartilage and the hypertrophic zone of the growth plate. Collecte five biological replications in three superficial, mid zone and deep zones of Articular Cartilage Assessed by Laser Captured Microdissection and Microarray(Superficial Zone vs Mid Zone vs Deep Zone)
Project description:This dataset was generated by RNA sequencing of bone marrow endothelial cells from femurs and tibiae of 6 day-old C57Bl/6 mice. Sample sets represent type H, type E and type L bone marrow endothelial cell subpopulations. Endothelial subpopulations are defined by their expression levels of Endomucin and CD31/Pecam1. Each dataset was generated in triplicates.
Project description:We have shown that removal of Lkb1 in chondorcytes results in enchondroma-like structure in postnatal mouse long bones. To furhter understand the role of Lkb1 in this process, we performed microarrrays to compare the transcriptional profile between control and conditional Lkb1 mutant (Col2a1-Cre; Lkb1c/c) chondrocytes. Postnatal day 30 mouse growth plate chondorcytes from control and mutant mouse femurs and tibiae were isolated for RNA extraction and hybridization on Affymetrix microarrays.
Project description:In progressed puberty, estrogen is responsible for the deceleration of growth by stimulating growth plate maturation. The mechanism of action is largely unknown. We obtained pubertal growth plate specimens of the same girl at Tanner stage B2 and Tanner stage B3, which allowed us to address this issue in more detail. Histological analysis showed that progression of puberty coincided with characteristic morphological changes associated with growth plate maturation, such as decreases in total growth plate height (p=0.002), height of the individual zones (p<0.001) and a increase in intercolumnar space (p<0.001). Microarray analysis of the specimens identified 394 genes (72% upregulated, 28% downregulated) changing with progression of puberty. Overall changes in gene expression were small (average 1.1 fold change). The 394 genes mapped to 13 significantly changing pathways (p<0.05) in majority belonging to extracellular matrix, cell cycle and cell death, which are all related to growth plate maturation. We next scanned the upstream promoter regions of the 394 genes for the presence of evolutionary conserved binding sites for transcription factors implemented in growth plate maturation such as Estrogen Receptor, Androgen Receptor, Elk1, Stat5b, CREBP and Runx2. Runx2 and Elk1, but not estrogen receptor binding sites were enriched and were present in 87 and 43 out of the 394 genes, respectively.In conclusion, our data suggest a role for Runx2 and Elk1 in growth plate maturation and provides suggestive evidence that the effect of estrogen on growth plate maturation is not mediated by activating genomic estrogen signalling in growth plate chondrocytes. 2 replicate samples for each developmental stage for a total of 6 samples
Project description:To initially determine changes on the transcriptome level that might account for the phenotypic observations in an unbiased fashion, we performed large-scale mRNA expression profiling 8 weeks after treatment with an AAV harboring a control or GAbpa-specific miRNA under the LP1 promoter. Mice were subjected to 16 h fasting or a 16 h fasting + 6 h refeeding cycle before termination of the experiment. All 8 AAV-injected experimental groups as NC and GAbpα knockdown samples for the different conditions (wt or db/db; fasting or refeeding) were included.