Lockhart, C, Scott, BR, Thoseby, B, and Dascombe, BJ. Acute effects of interset rest duration on physiological and perceptual responses to resistance exercise in hypoxia. J Strength Cond Res XX(X): 000-000, 2018-This study aimed to determine whether manipulating interset rest periods during resistance training in hypoxia impacts on physiological and perceptual responses to exercise. Twelve healthy males completed 1 repetition maximum (1RM) testing for the bilateral leg extension, before completing 4 separate randomized trials comprising 5 × 10 repetitions of leg extensions at 70% 1RM. Experimental trials were completed in both moderate hypoxia (FIO2 = 15%) and normoxia (FIO2 = 21%), using interset rest periods of both 60 and 180 seconds for each environmental condition. Near-infrared spectroscopy was used to quantify muscle oxygenation of vastus lateralis , and surface electromyography assessed the activation of vastus lateralis and medialis. Blood lactate concentration ([BLa]) and midthigh circumference were assessed before and immediately after each trial. Heart rate (HR) responses, blood oxygen saturation, and rating of perceived exertion (RPE) were also assessed after each set and the whole session RPE (sRPE). Perceived quadriceps soreness was reported before, immediately after, and at 24 and 48 hours after each trial. Muscle activation (sets 4-5), RPE (sets 3-5), and sRPE were significantly (p < 0.05) higher in the 60-second trials of the resistance exercise protocol. Significant increases (p < 0.01) were observed for [BLa] and midthigh circumference across sets within each condition. No significant main effect was observed for interset rest duration or environmental condition for muscle oxygenation, HR, or perceived quadriceps soreness. These findings indicate that performing resistance exercise in hypoxia or normoxia with shortened interset rest periods increases muscle activation and perceived exertion, without exacerbating muscle soreness.