Temperature is maintained in birds by skeletal muscle shivering as well as by non-shivering thermogenesis in a cold environment because they lack brown adipose tissue, which is a mammalian thermogenic organ. Chicks acquire cold tolerance after their skeletal muscles mature. Here, we found that muscle fibers transformed to the slow-twitch type with increasing gene expression of peroxisome proliferator-activated receptor-γ coactivator-1α (PGC-1α), and that the mass increased with decreasing myostatin gene expression, in the leg muscles of 7-day-old and younger chicks within 24 h of cold exposure. Muscle fibers did not transform and the mass did not increase within 24 h of cold exposure in muscles from chicks older than 8 days of age. Myostatin mRNA expression remained depressed in cold-tolerant muscles for 24 h, whereas cold-enhanced growth of the muscle continued for 48 h. Myostatin expression was depressed and muscle mass was increased only in chick leg muscles that comprised both fast- and slow-twitch fibers. These results suggest that the acute regulation of PGC-1αa and myostatin gene expression in leg muscles is required for chicks to acquire cold tolerance up to 7 days of age.