Project description: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.
Project description:To investigate differentially expressed lncRNAs in C2C12 myotubes with/without CoCl2 treatment, we used mouse lncRNA microarray to examine the expression of lncRNAs in C2C12 myotubes and C2C12 myotubes with CoCl2 treatment.
Project description:To investigate differentially expressed circRNAs in C2C12 myotubes with/without CoCl2 treatment, we used mouse circRNA microarray to examine the expression of circRNAs in C2C12 myotubes and C2C12 myotubes with CoCl2 treatment.
Project description:Introgressed variants from other species can be an important source of genetic variation because they may arise rapidly, can include multiple mutations on a single haplotype, and have often been pretested by selection in the species of origin. Although introgressed alleles are generally deleterious, several studies have reported introgression as the source of adaptive alleles-including the rodenticide-resistant variant of Vkorc1 that introgressed from Mus spretus into European populations of Mus musculus domesticus. Here, we conducted bidirectional genome scans to characterize introgressed regions into one wild population of M. spretus from Spain and three wild populations of M. m. domesticus from France, Germany, and Iran. Despite the fact that these species show considerable intrinsic postzygotic reproductive isolation, introgression was observed in all individuals, including in the M. musculus reference genome (GRCm38). Mus spretus individuals had a greater proportion of introgression compared with M. m. domesticus, and within M. m. domesticus, the proportion of introgression decreased with geographic distance from the area of sympatry. Introgression was observed on all autosomes for both species, but not on the X-chromosome in M. m. domesticus, consistent with known X-linked hybrid sterility and inviability genes that have been mapped to the M. spretus X-chromosome. Tract lengths were generally short with a few outliers of up to 2.7 Mb. Interestingly, the longest introgressed tracts were in olfactory receptor regions, and introgressed tracts were significantly enriched for olfactory receptor genes in both species, suggesting that introgression may be a source of functional novelty even between species with high barriers to gene flow.