The effects of sudden darkness on spontaneous motor activity in an open field and performance in an elevated plus-maze (EPM) were assessed in adult male and female rats. In the open field test, sudden darkness increased total locomotion, locomotion in central squares, rearing frequency (RF), and diminished defecation units (DU). In the dark, total locomotion remained elevated during the 20-min test period, while in the light total locomotion decreased significantly after the fourth minute, in both sexes. All the effects of sudden darkness in the open field test were more pronounced in female rats. In the EPM, sudden darkness increased the number of entries into the open arms, total entries, percentages of entries into the open arms, and time spent in the open arms. The changes were more significant in female than in male rats. These results show that sudden darkness increases general motor activity and suggest it diminishes habituation, fear, and anxiety. The results also suggest that this behavioral shift is sexually differentiated. Sudden darkness emerges as an experimental tool to simultaneously test physiologically-induced increases in spontaneous motor activity and decreases in anxiety.