Fig. 7: Photoactivation of aPVT^CRF-NAc projections suppresses food-seeking and induces avoidance behavior.

a (Top) Timeline of the cued food-seeking test during optogenetic activation of aPVT^CRF-NAc projections. (Bottom left) Representative micrograph showing the expression of Cre-dependent channelrhodopsin (AAV-DIO-ChR2-eYFP) under the control of AAV-CRF-Cre in aPVT. (Bottom left-center) Green areas represent the minimum (dark) and the maximum (light) viral expression in the aPVT. (Bottom right-center) Representative micrograph showing aPVT^CRF fibers in the NAc. (Bottom right) Purple dots represent the location of the optical fiber tips in the NAc. sm stria medullaris, 3V third ventricle, cc corpus callosum, MD mediodorsal thalamus, ac anterior commissure. b Schematic of the cued food-seeking test. c–e Optogenetic activation of aPVT^CRF-NAc projections (purple circles, n = 10) reduced c the number of lever presses (F_{(4, 72)} = 11.49, P < 0.001) and increased d the latency to press the lever (F_{(4, 72)} = 2.379, P = 0.059 with Bonferroni planned comparison test P = 0.012 for the first laser-on block and P = 0.054 for the second laser-on block), when compared to eYFP controls (gray circles, n = 10). No difference was found in e the percentage of time rats spent freezing during the illumination (F_{(1, 16)} = 4.316, P = 0.54). Blue shaded area represents laser-on trials (20 Hz, 5 ms pulse width,15 mW, 30 s duration). Each circle represents the average of two consecutive trials. f Schematic of the real-time place preference test. g–k Photoactivation of aPVT^CRF-NAc projections (purple bars, n = 9) reduced both g the percentage of time spent (F_{(1, 16)} = 18.73, P < 0.001) and h the distance traveled (F_{(1, 16)} = 16.78, P < 0.001) in the side of the chamber paired with laser stimulation (Side B), when compared to eYFP controls (gray bars, n = 9). i Representative tracks. No difference was found in locomotor activity measured as j total distance traveled (P = 0.711, t = 0.378) and (k) maximum speed (P = 0.212, t = 1.301) during the session. Blue shaded areas represent the sum of all laser-on epochs (20 Hz, 5 ms pulse width, 15 mW). c–e, g, h Two-way repeated-measure ANOVA followed by Bonferroni test. j, k Unpaired Student’s t test. Data are shown as mean ± SEM. *P < 0.05, ^#P between 0.05 and 0.099. See also Supplementary Movies 7 and 8.