Habitat loss and habitat fragmentation caused by anthropogenic activities are the main factors that constrain long-distance movement of ungulates. Mongolian gazelles (Procapra gutturosa) and Asiatic wild asses (Equus hemionus) in Mongolia are facing habitat fragmentation and loss. To better understand how their movements respond to potential anthropogenic and natural barriers, we tracked 24 Mongolian gazelles and 12 wild asses near the Ulaanbaatar-Beijing Railroad and the fenced international border between Mongolia and China between 2002 and 2012. None of the tracked gazelles crossed the railroad, even though gazelles were captured on both sides of the tracks at the start of the study. Similarly, we did not observe cross-border movements between Mongolia and China for either species, even though some animals used areas adjacent to the border. The both species used close areas to the anthropogenic barriers more frequently during winter than summer. These results suggest strong impacts by the artificial barriers. The construction of new railroads and roads to permit mining and other resource development therefore creates the threat of further habitat fragmentation, because the planned routes will divide the remaining non-fragmented habitats of the ungulates into smaller pieces. To conserve long-distance movement of the ungulates in this area, it will be necessary to remove or mitigate the barrier effects of the existing and planned roads and railroads and to adopt a landscape-level approach to allow access by ungulates to wide ranges throughout their distribution.