This study used a desktop virtual environmental simulation of 18 large-scale residential environments to test effects of plan layout complexity, physical differentiation, and gender on acquired spatial knowledge. One hundred sixty people (95 males and 65 females) were assigned at random to the different conditions. After a learning phase, participants were asked to (a) estimate the direction from the start point to a destination, (b) navigate to the destination, and (c) select the layout of the environment and sketch the route they took. Higher scores for acquired spatial knowledge emerged for simple over complex environments, for moderate or high differentiation over no differentiation, and for landmark or road differentiation over no such differentiation. For direction and navigation errors males performed marginally better than females, and for navigation errors younger respondents performed better. The results suggest that designs with physical differentiation and fewer choices at nodes will help people learn their way around.