Role of miR-1 in the mTORC1 signaling pathway in skeletal muscle

Abstract

Recent research advances in understanding the mechanisms and signaling pathways involved with the recovery process of muscle mass in senility, using different stimuli. These studies can contribute to the development of strategies to reduce or block the loss of muscle mass associated with aging. In this context, several studies have tried to investigate the signaling pathways and the role of Micro-RNAs involved in growth control, with maintenance of muscle phenotype as well as in understanding the process of recovery of muscle mass. This project aims to evaluate the role of microRNA (miR-1) in the regulation of hypertrophic signaling pathway (mTORC1) in skeletal muscle. Initially Bioinformatics tools will be used to identify possible targets of miR-1 in the mTORC1 pathway. After that, the genes will be validated targets of miR-1 by reporter gene construct 3'-UTR, with clone this region of the gene for luciferase. Each reporter gene is then co-transfected with miR-1 in fibroblast cells (3T3/NIH) and the activity of these genes is measured 48 hours later. For genes that show loss of expression is indicative that the 3'-UTR region contains a potential binding site miR-1. After this step, it is determined mRNA and protein expression of each target gene to determine if their pattern of expression is consistent with regulation of miRNA. This methodology will be held in Skeletal Muscle Biology Lab in the University of Kentucky, under the supervision of Prof.. Dr. John McCarthy. (AU)