Both signals were expected to be represented in the amygdala

as this brain region is known to play a crucial role in mediating vigilance and attention (Davis & Whalen, 2001), and in learning to predict aversive outcomes (Schiller et al., 2008; Eippert et al., 2012). Additionally, we tested for unsigned PE effects in the midbrain as recent buy Idelalisib animal studies suggest that the surprise-induced enhancement of learning depends on the integrity of midbrain–amygdala connections (Lee et al., 2006, 2008, 2010). Twenty-two healthy male subjects (mean age 26.9 years, range 21–33 years) participated in this study. Due to equipment malfunction one subject had to be excluded from further analysis. Only male volunteers were enrolled in the study to reduce variability in amygdala activation based on gender effects in conditioning and other emotional tasks (Milad et al., 2006; Cahill, 2010). The study was approved by the Ethics Committee of the Medical Board in Hamburg. All participants gave written informed consent

Copanlisib concentration and were paid for their participation. The paradigm used was a classical Pavlovian delay-conditioning procedure including an acquisition and a reversal phase. Reversal of cue–outcome associations provided a characteristic test to assess associability as the outcomes became surprising again with the beginning of the reversal phase (Holland & Gallagher, 1999; Li et al., 2011). Abstract fractal images served as visual CSs and the US was an electric shock (150 ms duration, 20 pulses/s of 0.01–100 mA) delivered to the dorsum of the right hand. Before the experiment, the intensity of the US was individually adjusted to be aversive. For this purpose, volunteers received shocks of varying intensity. They rated the aversiveness of the shocks on a rating scale ranging from 1 (not aversive) to 10 (very aversive and not tolerable). The intensity of the shock that received the rating score

9 (very aversive, but still bearable) was selected as the final intensity administered throughout the experiment. We used three different visual cues, each presented 40 times throughout the entire experiment. During Aprepitant acquisition, cue A co-terminated with a shock in 20/20 occasions (100% reinforcement, CS100), cue B was paired with shock in 10/20 occasions (50% reinforcement, CS50) and cue C was never followed by the US (CS–). Thus, the acquisition phase comprised the first 20 occurrences of each cue. In the reversal stage, the CS100 was not paired with shock (new CS–), whereas the CS50 was followed by a shock in every trial (new CS100) and the CS– was followed by a shock on 50% of the occasions (new CS50). The reversal phase immediately followed the acquisition phase. It started after a fixed number of trials such that the 61st trial was always the first trial of the reversal phase. Again, each cue was presented 20 times leading to a total experimental time of approximately 40 min.