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[Preprint]. 2024 Sep 24:2023.10.03.560731. Originally published 2023 Oct 3. [Version 2] doi: 10.1101/2023.10.03.560731

Figure 2: The BLA→DMS pathway is typically activated by rewards during action-outcome learning, chronic stress attenuates this and instead progressively recruits CeA→DMS pathway activity to learning.

Figure 2:

(a) Intersectional approach for fiber photometry calcium imaging of DMS-projecting BLA or CeA neurons. (b) Schematic representation of retrograde AAV-cre in DMS and cre-dependent GCaMP8s expression and optical fiber tips in BLA or CeA for all subjects. (c) Procedure schematic. Stress, chronic unpredictable stress. Lever presses earned food pellet rewards on a random-ratio (RR) reinforcement schedule. (d-i) Fiber photometry recordings of GCaMP8s in BLA→DMS neurons during instrumental lever press – food-pellet reward learning. (d) Representative images of retro-cre expression in DMS and immunofluorescent staining of cre-dependent GCaMP8s expression and fiber placement in BLA. (e) Press rate across training. Training: F(1.72, 32.66) = 81.40, P < 0.0001. (f-g) Z-scored Δf/F BLA→DMS GCaMP8s fluorescence changes aligned to bout-initiating presses (f) and reward collection (g) averaged across trials for each subject (shading reflects between-subject s.e.m.) across each training session. (h-i) Quantification of area under the BLA→DMS GCaMP8s Z-scored ∆f/F curve (AUC) during the 3-s period prior to initiating presses (h; Training: F(2.49, 47.38) = 0.91, P = 0.43) or following reward collection (i; Stress: F(1, 19) = 24.13, P < 0.0001). Control N = 9 (4 male), Stress N = 12 (5 male). (j-o) Fiber photometry recordings of GCaMP8s in CeA→DMS neurons during instrumental lever press – food-pellet reward learning. (j) Representative immunofluorescent image of retro-cre expression in DMS and cre-dependent GCaMP8s expression and fiber placement in CeA. (k) Press rate across training. Training: F(1.51, 30.23) = 65.61, P < 0.0001. (l-m) Z-scored Δf/F CeA→DMS GCaMP8s fluorescence changes aligned to bout-initiating presses (l) and reward collection (m) averaged across trials for each subject across each training session. (n-o) Quantification of CeA→DMS GCaMP8s Z-scored ∆f/F AUC during the 3-s period prior to initiating presses (n; P = 0.58; Stress: F(1, 20) = 0.74, P = 0.40) or following reward collection (o; Training x Stress: F(3, 60) = 4.51, P = 0.006). Control N = 11 (6 male), Stress N = 11 (4 male). Males = closed circles/solid lines, Females = open circles/dashed lines. *P <0.05, **P <0.01, ***P < 0.001.