Ogasawara R, Sato K, Matsutani K, Nakazato K, Fujita S. The order of concurrent endurance and resistance exercise modifies mTOR signaling and protein synthesis in rat skeletal muscle. Am J Physiol Endocrinol Metab 306: E1155-E1162, 2014. First published April 1, 2014; doi:10.1152/ajpendo.00647.2013.-Concurrent training, a combination of endurance (EE) and resistance exercise (RE) performed in succession, may compromise the muscle hypertrophic adaptations induced by RE alone. However, little is known about the molecular signaling interactions underlying the changes in skeletal muscle adaptation during concurrent training. Here, we used an animal model to investigate whether EE before or after RE affects the molecular signaling associated with muscle protein synthesis, specifically the interaction between RE-induced mammalian target of rapamycin complex 1 (mTORC1) signaling and EE-induced AMP-activated protein kinase (AMPK) signaling. Male Sprague-Dawley rats were divided into five groups: an EE group (treadmill, 25 m/min, 60 min), an RE group (maximum isometric contraction via percutaneous electrical stimulation for 3 ϫ 10 s, 5 sets), an EE before RE group, an EE after RE group, and a nonexercise control group. Phosphorylation of p70S6K, a marker of mTORC1 activity, was significantly increased 3 h after RE in both the EE before RE and EE after RE groups, but the increase was smaller in latter. Furthermore, protein synthesis was greatly increased 6 h after RE in the EE before RE group. Increases in the phosphorylation of AMPK and Raptor were observed only in the EE after RE group. Akt and mTOR phosphorylation were increased in both groups, with no between-group differences. Our results suggest that the last bout of exercise dictates the molecular responses and that mTORC1 signaling induced by any prior bout of RE may be downregulated by a subsequent bout of EE. concurrent exercise; mammalian target of rapamycin complex 1; adenosine 5=-monophosphate-activated protein kinase; resistance exercise; endurance exercise SKELETAL MUSCLE IS A HIGHLY PLASTIC TISSUE that adapts differently to different stimuli, including various types of muscle contractions. Chronic muscle contraction induces diverse metabolic and morphological adaptations because of the cumulative effects of repeated bouts of exercise, and certain molecular and cellular responses lead to specific adaptations (3, 10, 47). The well-known adaptive features of chronic muscle contraction include muscle hypertrophy and increased strength. These features are induced by high-intensity muscle contraction, such as that involved in resistance exercises (RE) (50, 52). Other typical adaptations are increases in oxidative capacity and glucose uptake, which are induced by low-intensity, highvolume muscle contraction, such as that observed during endurance exercises (EE) (5, 28, 53).Concurrent training, a combination of EE and RE in succession, is used widely in exercise prescription to simultaneously improve muscular strength and cardiovascular function. Interestingly, ac...