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Previous research has shown that low central serotonin, induced by acute tryptophan depletion (ATD), results in depressed mood and impairs cognition in healthy volunteers with a predisposition for depression. It remains unknown whether ATD affects emotional processing via mood changes or directly. In the present study we investigated the interaction between vulnerability for depression and the effect of ATD on mood, cognition and the associated brain activation. In a previous functional MRI study, we tested the effect of ATD during a combined cognitive and emotional Stroop task in healthy women without a family history of depression (FH-). In this study, we present the data of an additional group of 12 healthy women with a positive family history of unipolar depression (FH+). The effect of ATD on mood and Stroop performance was different for the FH+ group as compared with the FH- group. Scores on the depression sub-scale of the Profile of Mood States (POMS) did not correlate with performance changes, but did correlate with the anterior cingulate cortex response during Stroop interference. This study showed that a family history of unipolar depression interacts with the effect of ATD.