A. Preparation for opto-Pavlovian task combined with dLight recordings in the NAc. Scale bar; 1mm. White dashed lines indicate fiber tracts. B. Schematic for opto-Pavlovian task. One cue was associated with the laser delivery while the other cue was not. C. dLight recordings in the NAc of a representative mouse around the laser cue presentation at session (left) and grouped data (middle). dLight recordings of non laser trials are also shown (right). D. Area under the curve (AUC) of dLight signal in the NAc around the cue presentations (0-1.5 seconds) across sessions. E. AUC at session 1 and 10 for the non-laser cue trials and laser cue trials. AUC of non-laser cue trials were unchanged while AUC around laser cue trials showed increase (bottom). One-way analysis of variance; F (3, 12) = 5.927, P=0.0101. Tekey’s multiple comparison test. Session 1 non-laser trials vs. session 10 non laser trials ; P=0.9256. Session 1 laser trials vs. session 10 laser trials ; P=0.0196.

A. Schematic for the omission sessions. Two thirds of laser associated cue was followed by the laser stimulation while the other one third of the laser associated cue failed to trigger the laser stimulation. B.dLight recordings of a representative mouse during omission sessions. dLight signal around the laser cue presentation is shown here. White asterisks indicate omission trials, while in the other trials, the laser stimulation was delivered. C. dLight recordings in the NAc during omission trials. A dip of dLight signals was observed. One sample t test; t=4.176, df=3.P= 0.0250.

A. Schematic for the dLight recording in the LH while stimulating dopamine neurons in the VTA (left). Coronal image of the LH of a mouse infected with AAV-hSyn-DIO-Chrimson-tdTomato in the VTA and AAV-hSyn-dLight1.3b in the LH (right). White dashed lines indicate fiber tracts. Scale bar; 1mm. B. dLight signal in the LH during dopaminergic stimulation in the VTA at several number of pulses (20Hz, 10ms duration for each pulse). C. dLight recordings during the laser cue presentation of a representative mouse at session 1. D. dLight recordings around the laser cue presentation (left) and non-laser cue presentation (right) at session 1. E. dLight recordings during the laser cue presentation of a representative mouse at session 10. F. dLight recordings around the laser cue presentation (left) and non-laser cue presentation (right) at session 10. G. Area under the curve (AUC) of dLight signal in the LH around the cue presentations (0-1.5 seconds) across sessions. AUC of non-laser cue trials were unchanged while AUC around laser cue trials showed increase (right). One-way analysis of variance; F (3, 12) = 4.774, P=0.0205. Tekey’s multiple comparison test. Session 1 non-laser trials vs. session 10 non laser trials ; P= 0.7925. Session 1 laser trials vs. session 10 laser trials ; P= 0.0339. H. dLight recordings in the LH during omission trials. A dip of dLight signals was observed. One sample t test; t=3.193, df=3.P= 0.0496.

A. Schematic of the preparation for opto-Pavlovian task combined with orexin promoter GCaMP recordings in the LH. B. Coronal image of a mouse brain slice infected with AAV-hSyn-DIO-ChrimsonR-tdTomato in the VTA and AAV1-hOX-GCaMP6S in the LH (left. Scale Bar; 1mm). White dashed lines indicate fiber tracts. Zoom of infected LH with AAV1-hOX-GCaMP6s and co-localization orexin IR and GCaMP6s (right. Scale Bars; 50 μm). C. Orexin promoter GCaMP recordings in the LH of a representative mouse around the laser cue presentation at session 1 (left), grouped data (middle) and recordings during non laser trial (right). D. Orexin promoter GCaMP recordings in the LH of a representative mouse around the laser cue presentation at session 10 (left), grouped data (middle) and recordings during non laser trial (right). E. Area under the curve (AUC) of orexin promoter GCaMP signal in the LH around the cue presentations (0-1.5 seconds) across sessions. One-way analysis of variance; F (3, 12) = 9.904, P=0.0014. Tekey’s multiple comparison test. Session 1 non-laser trials vs. session 10 non laser trials ; P= 0.4323. Session 1 laser trials vs. session 10 laser trials ; P= 0.0024. G. Schematic for the omission sessions. F. Orexin promoter GCaMP recordings during stimulation trials (left) and omission trials (middle and right). AUC around the omission was higher than zero. One sample t test; t=4.693, df=3.P= 0.0183.

A. Schematic for the orexin promoter GCaMP recording in the LH while stimulating dopamine terminals in the LH. B. Orexin promoter GCaMP signals of a representative mouse. Recordings were performed while mice were freely moving (top) and anesthetized with isoflurane (bottom). Red bars indicate the stimulation. (20Hz, 100 pulses, 10 ms duration). C. Orexin promoter GCaMP signals around the stimulation of dopamine terminals in the LH while animals were freely moving (left) and anesthetized (right). D. AUC at 0 to 20 seconds was not significantly different between freely moving and anesthetized conditions. Paired t test, t=1.923 df=2.P= 0.1944. E. In freely moving condition, recordings were performed after mice received the intraperitoneal injection of vehicle (left), SCH 23390 (1mg/kg, middle), and raclopride (1mg/kg, right). F. Area under the curve (AUC) at 0-5 seconds. Black line indicates the mean for each condition and grey lines show individual mice. The administration of raclopride decreased the AUC significantly while SCH 23390 did not change the AUC. One-way analysis of variance; F (3, 6) = 5.305, P=0.04. Tekey’s multiple comparison test. vehicle vs. SCH 23390; P= 0.8145. vehicle vs. raclopride; P= 0.0476.