Rats with lesions of the medial septum were more likely to begin swimming in the wrong direction, swim farther, and, therefore, require more time to find a platform hidden in a Morris water tank than were control rats. Although the performance of the rats with medial septal lesions did improve over trials, their asymptotic performance also failed to equal that of the controls. Movement of the platform to a new position in the tank disrupted the performance of both groups, and, again, the rats with medial septal lesions were slower to locate the moved platform. However, this deficit was completely eliminated when a visual cue indicating the location of the moved platform was introduced. We suggest that these data indicate that damage to the septo-hippocampal cholinergic projection system produces a deficit in the formation or utilization of a spatial map (reference memory) that represents the location of a place with respect to the surrounding distal cues.