Introduction: The maintenance of skeletal muscle mass and function is critical for health and wellbeing, with deterioration in muscle size and strength associated with numerous chronic diseases. Skeletal muscle protein balance is highly dependent on the activity of the mammalian target of rapamycin complex 1 (mTOR) a serine/threonine protein kinase, which when active stimulates protein synthesis and offsets protein degradation. Mechanism(s) of mTOR activation in human skeletal muscle are still not fully understood. Cell and rodent based studies have reported mTOR to translocate to specific intracellular areas (e.g. the lysosome) in response to anabolic signals such as amino acids, growth factors and resistance-type exercise. To date, however, whether mTOR translocates in human skeletal muscle in response to exercise and nutrition is unknown, as are the associated proteins involved in mTOR activation and repression.Methods: In the present study, 14 young healthy male volunteers (24.7 ± 2.7 yrs, 177.4±6.8 cm, BMI 26.1±2.2 kg m−2) performed an acute bout of resistance exercise (4 sets of leg press and knee extension at 8-10 repetition maximum) prior to ingesting a beverage providing 20/44/1g of protein/carbohydrate/fat (PRO: n=7) to enhance muscle protein synthesis and anabolic signalling or an energy-free control (CON: n=7). Muscle biopsies were collected from the vastus lateralis pre and post-exercise and 1h and 3h after beverage ingestion. Skeletal muscle samples were immediately mounted in optimal cutting temperature medium and frozen in isopentane cooled by liquid nitrogen. Samples were serially sectioned for subsequent immunofluorescent analysis. Results: mTOR and LAMP2 were found to be highly colocalised in human skeletal muscle in both basal and exercise stimulated conditions (Pearson’s correlation coefficient: 0.45±0.02 for pre, 0.40±0.03 0h, 0.42±0.02 1h and 0.44±0.02 3h post exercise). In both CON and PRO we observed mTOR translocation to the sarcolemmal membrane. Quantification of mTOR translocation (Pearson’s correlation coefficient) found that PRO resulted in a greater duration of mTOR translocation to the sarcolemmal membrane (1 and 3h) post exercise (0.23±0.01 pre, 0.26±0.02 1h, p<0.05 and 0.25±0.008 3h post exercise respectively, p<0.05), while significant sarcolemmal translocation of mTOR was only found 1h after resistance exercise in the CON group (0.24±0.02 pre, 0.27±0.02, 1h post exercise, p<0.01). Conclusion: We present for the first time that mTOR/LAMP2 complexes are highly co-localized in human skeletal muscle. Moreover, in response to an anabolic stimulus, these complexes translocate to the sarcolemmal membrane with this event prolonged by nutrient ingestion known to enhance rates of muscle protein synthesis and mTOR signalling.