Calpastatin (CAST) is an endogenous calpain inhibitor and its main function is to modulate the proteolytic action of enzymes responsible for post-mortem myofibril deterioration. The myostatin gene (GDF-8) acts as a negative regulator of skeletal muscle growth. The expression of these two genes, as well as their interaction, affects the quality of the meat, especially the tenderness phenotype. We evaluated the genetic groups Santa Inês, ½ Dorper-Santa Inês and ½ White Dorper-Santa Inês, slaughtered with 2.0 mm, 2.5 mm and 3.0 mm of fat thickness, comparing the levels of expression of the CAST and GDF-8 genes with the weight performance and carcass traits, especially the shear force values. We found significantly higher expression of myostatin and calpastatin in the Santa Inês genetic group. The ½ Dorper-Santa Inês genetic group had the lowest expression of these genes when slaughtered with 2.0 and 2.5 mm of fat thickness. In conclusion, the Santa Inês breed had the lowest phenotype values for meat tenderness, and the ½ Dorper-Santa Inês breed had the best performance for this characteristic. We suggest that high levels of the expression of the CAST and GDF-8 genes are associated with lower values of lamb meat tenderness, and that tenderness is related to the stage of muscular growth and development.