We investigated metabolic costs, muscle activity, and perceptual responses during forward and backward running at matched speeds at different body weight support (BWS) conditions. Participants ran forward and backward on a lower body positive pressure treadmill at 0%BWS, 20%BWS, and 50%BWS conditions. We measured oxygen uptake, carbon dioxide production, heart rate, muscle activity, and stride frequency. Additionally, we calculated metabolic cost of transport. Furthermore, we used rating of perceived exertion and feeling scale to investigate perceptual responses. Feeling scale during running was higher with increasing BWS (0–50%BWS), regardless of running direction (p < 0.05). Oxygen uptake, heart rate, and metabolic cost of transport were influenced by the interaction of running direction and BWS (p < 0.01). For example, metabolic cost of transport during backward running was greater than when running forward only when running at 0%BWS (i.e., 4.4 ± 1.1 and 5.8 ± 1.4 J/kg/m for forward and backward running, respectively: p < 0.001). However, rectus femoris muscle activity, stride frequency, and rating of perceived exertion during backward running were averages of 113.5%, 11.3%, and 2.8 rankings greater than when running forward, respectively, regardless of BWS (p < 0.001). We interpret our observations to indicate that environment (in the context of effective body weight) is a critical factor that determines self-selected movement patterns during forward and backward running.