AbstractSerotonin transporter gene variance has long been considered an important factor contributing to depression. However, meta-analyses yielded inconsistent findings recently, asking for further understanding of the link between the gene and depression-related symptoms. One key feature of depression is anhedonia. Anhedonia is not one united feature but involves consummatory, anticipatory, and decisional aspects. These different aspects of anhedonia can be particularly well investigated in rodents. While data exist on the effect of serotonin transporter gene knockout in rodents on consummatory and anticipatory anhedonia, with mixed outcomes, the effect on decisional anhedonia has not been investigated thus far in rats. Here, we tested whether serotonin transporter knockout (5-HTT−/−) contributes to decisional anhedonia. To this end, we established a novel touchscreen-based task employing a ‘go/go’ task for decision-making under ambiguity. Both genotypes of rats were able to reach 70% accuracy across three consecutive sessions. However, 5-HTT+/+ rats performed more optimal decision-making compared to 5-HTT−/− rats during the course of discriminative conditioning for the conditioned stimulus associated with high reward value at the beginning. 5-HTT−/− rats gradually reduced response time to generalization stimuli compared to 5-HTT+/+ rats across the three generalization test sessions without altering the monotonic graded responses towards the stimuli. These data indicate that 5-HTT−/− rats initially processed the reward related ambiguous information in an anhedonic mode, which gradually over sessions turned into a hedonic mode of processing. The results suggest that the association between the serotonin transporter gene and anhedonia may dynamically change over time when processing ambiguous information. This may explain the inconsistent findings on the association between the serotonin transporter gene and various aspects of depression in humans.