Multi-user Equipament approved in grant 2019/11820-5: ChemiDoc Imaging System and accessories

Abstract

Recently, the World Health Organization classified sarcopenia as a disease, which will allow the establishment of universal clinical guidelines for its diagnosis and treatment. In this sense, the rev-erb-alpha protein is a promising molecular target for the prevention and treatment of sarcopenia, since it plays an essential role in the regulation of autophagy, mitochondrial content, oxidative function, and skeletal muscle mass maintenance. Therefore, the present thematic project will investigate the function of the gene nuclear receptor subfamily 1 group D member 1 (Nr1d1) in the development of sarcopenia associated with aging through experiments on cells, rodents, and humans. For this, the following subprojects will be used: 1) Effect of aging on Nr1d1 and its relationship with sarcopenia; 2) Effect of global or conditional deletion of Nr1d1 on aging; 3) Interactome analysis of rev-erb-alpha protein through immunoprecipitation of the target protein and identification of possible ligands by mass spectrometry in cell culture; 4) Effects of pharmacological, nutritional and genetic activation of Nr1d1 on the aging process; 5) Effects of physical exercise on the aging-induced sarcopenia: the role of rev-erb-alpha in the skeletal muscle. The methodologies used in this project will be RNA-seq, immunoblotting, Real time-RTqPCR, interactome, mass spectrometry, gene therapy (CRISPR/Cas9 and adeno-associated virus vector), histology, immunohistochemistry, immunofluorescence, bioinformatics, and lysosomal autophagy flow. According to the statistical distribution of the data, parametric or non-parametric tests will be used to compare the studied parameters between the experimental groups. (AU)