A fundamental cognitive function in humans and other species is the ability to orient and navigate in space. Using behavioral and brain imaging methods to investigate the processing of visual cues mediating spatial navigation and orientation in humans, I examined the cognitive and neural representations of two distinct classes of visual cues in spatial cognition: directional cues and positional cues. Directional (compass) cues include gradient information, (e.g., geographic slant), distal objects and geometric cues, and provide primarily directional information. Positional cues are proximate and unique objects, whose aspect changes rapidly with the movement of the observer, and provide relatively more precise positional information within the local object array. We present here a series of studies that examined sex differences, neural correlates, and developmental trajectories of the processing of these two classes of cues. In part one of this thesis, we present two behavioral studies on sex differences. Our results suggest that males compared to females rely more on directional cues during spatial tasks. In chapter 3, we demonstrated that navigation under directional cues was more accurate in males. In chapter 4, we showed that adding an additional directional cue to the environment improved male performance in a spatial memory task, and removing the directional cues impaired males more than females in a navigation task. The evidence for the female advantage in positional cue processing, however, was not robust. In the second part of this thesis, we investigated the neural correlates of spatial navigation with directional and positional cues. We found evidence for different neural signatures of the two cue types. Positional cues had greater activation in the posterior hippocampus whereas directional cues had greater activation in the anterior hippocampus. In the third part of this thesis, we investigated the developmental trajectories of the encoding of the two types of cues in 2-dimensional (2D) natural scenes. Our findings suggest the representation of positional cue goes through a prolonged maturation process compared to directional cues, and has distinct neural signatures. Taken together, these studies provide converging evidence in humans for the dissociable, parallel systems supporting the representations of directional and positional cues.