Unknown,Transcriptomics,Genomics,Proteomics

Dataset Information

PPARg dependent expression changes during differentiation of 3T3L1 cell into adipocytes

ABSTRACT: We profiled PPARg dependent gene expression changes during differntiation of 3T3L1 cell using PPARg siRNA 3T3-L1 (Pre-adipocyte) cell line was induced to differentiate using standard adipocyte differentiation media (IBMX, Dex and Insulin) 48hrs post-confluency. RNA was harvested at day -2 (confluent fibroblasts), 48hrs post-induction with IBMX, DEX and Insulin (day=0) and for each subsequent day after rosiglitazone treatment. Illumina beadchip microarrays were used to determine expression profiles of genes differentially regulated in cells transfected with either siRNA targeting PPARgamma or a non-targeting control siRNA. 3T3L1 cell were induced to differentiate into adipocytes using IBMX, DEX and Insulin. RNA from cell treated with PPARg-specific siRNA and non-specific siRNA was isolated at different timepoints. Illumina MouseRef-8 v1.1 Bead chips were used for expression profiling

ORGANISM(S): Mus musculus

SUBMITTER: Mohamed Hamza

PROVIDER: E-GEOD-12929 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

De-novo identification of PPARgamma/RXR binding sites and direct targets during adipogenesis.

Hamza Mohamed Sabry MS Pott Sebastian S Vega Vinsensius B VB Thomsen Jane S JS Kandhadayar Gopalan Srinivasan GS Ng Patrick Wei Pern PW Chiu Kuo Ping KP Pettersson Sven S Wei Chia Lin CL Ruan Yijun Y Liu Edison T ET

PloS one 20090320 3

<h4>Background</h4>The pathophysiology of obesity and type 2 diabetes mellitus is associated with abnormalities in endocrine signaling in adipose tissue and one of the key signaling affectors operative in these disorders is the nuclear hormone transcription factor peroxisome proliferator-activated receptor-gamma (PPARgamma). PPARgamma has pleiotropic functions affecting a wide range of fundamental biological processes including the regulation of genes that modulate insulin sensitivity, adipocyte ...[more]

PMID: 19300518

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Project description:Differentiation of 3T3-L1 preadipocytes can be induced by a 2-d treatment with a factor "cocktail" (DIM) containing the synthetic glucocorticoid dexamethasone (dex), insulin, the phosphodiesterase inhibitor methylisobutylxanthine (IBMX) and fetal bovine serum (FBS). We temporally uncoupled the activities of the four DIM components and found that treatment with dex for 48 h followed by IBMX treatment for 48 h was sufficient for adipogenesis, whereas treatment with IBMX followed by dex failed to induce significant differentiation. Similar results were obtained with C3H10T1/2 and primary mesenchymal stem cells. The 3T3-L1 adipocytes differentiated by sequential treatment with dex and IBMX displayed insulin sensitivity equivalent to DIM adipocytes, but had lower sensitivity to isoproterenol-stimulated lipolysis and reduced triglyceride content. The nondifferentiating IBMX–then-dex treatment produced transient expression of adipogenic transcriptional regulatory factors C/EBP{beta} and C/EBP{delta}, and little induction of terminal differentiation factors C/EBP{alpha} and PPAR{gamma}. Moreover, the adipogenesis inhibitor preadipocyte factor-1 (Pref-1) was repressed by DIM or by dex-then-IBMX, but not by IBMX-then-dex treatment. We conclude that glucocorticoids drive preadipocytes to a novel intermediate cellular state, the dex-primed preadipocyte, during adipogenesis in cell culture, and that Pref-1 repression may be a cell fate determinant in preadipocytes. We sought to define the genes differentially expressed in response to dexamethasone (dex) in 3T3L1 mouse preadipocytes. We treated 3T3L1 cells with dex or DMSO vehicle for 2 or 36 hrs and then prepared total RNA. We hybridized each sample (Cy5) to a pool of the samples, which served as the reference (Cy3) channel. All linear models for differential gene expression were constructed with the "limma" package (Smyth, 2004) in BioConductor (Gentleman et al., 2004). A linear model for dex was constructed to compare the 4 biological replicates of DMSO vehicle treatment with dex treatment at either 2 or 36hr. Genes were considered differentially expressed if at least one probe had log-odds greater than 50:50 in the linear model.

Dataset Information

PPARg dependent expression changes during differentiation of 3T3L1 cell into adipocytes

Publications

De-novo identification of PPARgamma/RXR binding sites and direct targets during adipogenesis.

Similar Datasets

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