Transcriptomics

Dataset Information

15

Transcription profiling of mouse C2C12 myotubes treated overnight with 500 uM palmitate or 1% bovine serum albumin control to identify mediators of obesity-linked reductions in PGC-1


ABSTRACT: To identify mediators of obesity-linked reductions in PGC-1, we tested the effects of cellular nutrients in C2C12 myotubes. While overnight exposure to high insulin, glucose, glucosamine, or amino acids had no effect, saturated fatty acids (FA) potently reduced PGC-1a and b mRNA expression. Experiment Overall Design: Cell culture - Mouse C2C12 myoblasts (ATCC, Manassas, VA) were maintained in Dulbecco's modified Eagle's medium (DMEM) supplemented with 20% fetal bovine serum (Invitrogen), at a confluency of 60-70%. To initiate differentiation, cells were allowed to reach 100% confluency and medium was changed to DMEM containing 2% horse serum (Invitrogen) and changed every 2 days. Full differentiation, with myotube fusion and spontaneous twitching, was observed at 5 days. Fatty acid stock preparation - Fatty acids were dissolved in 0.1 N sodium hydroxide (final concentration 100 mM) at 65 degrees C for 2 hours and then complexed with 10% fatty acid-free BSA, yielding a final stock of 5 mM. Three replicates for each fatty-acid. Microarray analysis - RNA was isolated as described from C2C12 myotubes treated overnight with 500 uM palmitate or 1% BSA, and cRNA was synthesized. 10 mg of cRNA were hybridized to Affymetrix mouse 430A 2.0 arrays. Intensity values were quantified using MAS 5.0 software. MAPPFinder (www.genmapp.org) was used to integrate expression data with known pathways. Transcription profiling of mouse C2C12 myotubes treated overnight with 500 uM palmitate or 1% BSA to identify mediators of obesity-linked reductions in PGC-1.

REANALYSED by: E-GEOD-6766

INSTRUMENT(S): 418 [Affymetrix]

ORGANISM(S): Mus musculus  

SUBMITTER: Sarah Crunkhorn  

PROVIDER: E-GEOD-6766 | ArrayExpress | 2007-11-15

SECONDARY ACCESSION(S): GDS2648GSE6766PRJNA98939

REPOSITORIES: GEO, ArrayExpress

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Publications

Peroxisome proliferator activator receptor gamma coactivator-1 expression is reduced in obesity: potential pathogenic role of saturated fatty acids and p38 mitogen-activated protein kinase activation.

Crunkhorn Sarah S   Dearie Farrell F   Mantzoros Christos C   Gami Hiral H   da Silva Wagner S WS   Espinoza Daniel D   Faucette Ryan R   Barry Kristen K   Bianco Antonio C AC   Patti Mary Elizabeth ME  

The Journal of biological chemistry 20070406 21


Peroxisome proliferator activator receptor-gamma coactivator 1 (PGC-1) is a major candidate gene for diabetes-related metabolic phenotypes, contributing to decreased expression of nuclear-encoded mitochondrial genes in muscle and adipose tissue. We have demonstrated that muscle expression of PGC-1alpha and -beta is reduced in both genetic (Lep(ob)/Lep(ob)) and acquired obesity (high fat diet). In C57BL6 mice, muscle PGC-1alpha expression decreased by 43% (p < 0.02) after 1 week of a high fat die  ...[more]

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