In two experiments rats were required to escape from a circular pool by swimming to an invisible platform that was located in the same place relative to one configuration of two landmarks (X and Y). The two landmarks were placed relatively far and equidistant from the hidden platform. Training could be either on consecutive days (Experiment 1) or every fourth day (Experiment 2). Subsequent test trials, without the platform, revealed a preference for searching in the correct quadrant of the pool. In Experiment 1 such a test performance was identical in two groups of females, one tested with high hormonal levels (i.e., in the proestrus phase) and the second one tested with low hormonal levels (i.e., either in the estrus, metaestrus or diestrus phase); in addition, these two groups differed from a third group of male rats (i.e., males had a better performance than females). Experiment 2 replicated the females’ previous results with a better procedure. The experiment compared the performance of two groups of female rats which were both trained and tested always in the same estrus phase, one group in the proestrus phase, and the second group in the estrus phase. The implication of these results is that the estrus cycle has little impact on the performance of female rats when landmark learning in a navigation task.