RNA-Seq transcriptome profiling of soleus from adult and aged mice following disuse muscle atrophy and recovery. Mus musculus

Rodent hind limb unloading was used as a model for reduced muscle activity and eventual atrophy. After a 10 day period of unloading, mice in this study were “reloaded” for 3 days and regained use of their hind limbs. We report the application of Next-generation sequencing (NGS) technology for high-throughput profiling of mRNA in soleus muscle of adult (6 mo) and aged (22-24 mo) mice. Our goal was to determine the effects of hind limb unloading and reloading on mRNA profiles in soleus muscle and compare between adult and aged mice. We find that there are distinct response in the profile of fatty acid oxidation, TCA cycle, ETC oxidative phosphorylation gene expression patterns in response to unloading and reloading. The repsonses are generally simialr between young and old mice. Overall design: RNA-Seq was performed with total RNA isolated from snap frozen soleus muscle. Male C57BL/6J mice (Adult 6 mo and Old 22-24 mo) were obtained from the NIA's Aged Rodent Colony Charles River Laboratories, Madison, WI) and were subjected to the following experimental conditions: 1) hind limb unloaded for 10 days, 2) hind limb unloaded for 10 days and reloaded for 3 days, or 3) maintained under control conditions for 10 days. Mice were fed ad libitum with a standard chow diet (#2916, Harlan-Teklad, Houston, Tx) and water/hydrogel (ClearH2O, Westbrook, ME, US) and housed at 22C with a 12-h light/dark cycle. All soleus samples were harvested following a 4 hour fast and mice were sacrificed at ~11am by CO2 asphyxiation and cervical dislocation.