Abstract
Exercise has beneficial effects on human health and is affected by two different pathways; motoneuron and endocrine. For the advancement of exercise research, in vitro exercise models are essential. We established two in vitro exercise models using C2C12 myotubes; EPS (electrical pulse stimulation) for a motoneuron model and clenbuterol, a specific β2 adrenergic receptor agonist, treatment for an endocrine model. For clenbuterol treatment, we found that Ppargc1a was induced only in low glucose media (1 mg/mL) using a 1-h treatment of 30 ng/mL clenbuterol. Global transcriptional changes of clenbuterol treatment were analyzed by RNA-seq and gene ontology analyses and indicated that mitogenesis and the PI3K-Akt pathway were enhanced, which is consistent with the effects of exercise. Cxcl1 and Cxcl5 were identified as candidate myokines induced by adrenaline. As for the EPS model, we compared 1 Hz of 1-pulse EPS and 1 Hz of 10-pulse EPS for 24 h and determined Myh gene expressions. Ten-pulse EPS induced higher Myh2 and Myh7 expression. Global transcriptional changes of 10-pulse EPS were also analyzed using RNA-seq, and gene ontology analyses indicated that CaMK signaling and hypertrophy pathways were enhanced, which is also consistent with the effects of exercise. In this paper, we provided two transcriptome results of in vitro exercise models and these databases will contribute to advances in exercise research.
Highlights
Exercise has beneficial effects in the prevention and treatment of a number of diseases, such as metabolic diseases, psychiatric diseases, neurological diseases, cardiovascular diseases, musculo-skeletal disorders, and cancer [1,2,3,4]
Chronic reactive genes are not induced by a single bout of exercise; once the gene expression is modified, it is maintained for longer period of time, such as with Myh genes [11]
Acute transcriptional induction by exercise was observed at one hour after exercise [25,26]; the duration of clenbuterol treatment was determined to be one hour
Summary
Exercise has beneficial effects in the prevention and treatment of a number of diseases, such as metabolic diseases, psychiatric diseases, neurological diseases, cardiovascular diseases, musculo-skeletal disorders, and cancer [1,2,3,4]. Motoneuron firing induces intramyofibrillar Ca2+ release from the sarcoplasmic reticulum to trigger muscle contraction through myosin heavy chain ATPase activity, while systemic conditions, such as increased sympathetic nervous system activity, release of fatty acids, and glucocorticoid secretion, are changed. These stimuli synergistically or additively induce transcriptomic activation or repression in skeletal muscles [7,8,9]. CaMK phosphorylates HDAC (histone deacetylase) to induce nuclear export, which complexes with MEF2 proteins to repress MEF2-dependent genes, such as Myh7 Both calcium-induced transcriptions lead fiber-type shifts to a slow type [12]. Global transcriptional analyses of 1 h of clenbuterol treatment were performed as a model of endocrine stimulation
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