Extended Data Fig. 2: Plasticity in DA release and DAN activity dynamics.

a, Behavioural parameters demonstrating that mice are able to learn the visual-cue-guided operant conditioning described in Fig. 2a. Top: from the left, success rate (number of rewarded trials/total number of trials, one-way repeated-measures ANOVA, F(6.298, 384.2) = 107.8, P < 0.0001), entering failure rate (number of receptacle zone entering failure trials/total number of trials, one-way repeated-measures ANOVA, F(5.782, 352.7) = 103.8, P < 0.0001), occupancy failure rate (number of premature receptacle zone exit trials/number of receptacle zone entering success trials, one-way repeated-measures ANOVA, F(4.253, 225.4) = 7.324, P < 0.0001), time spent in zone (time spent in a zone/total session time, receptacle: one-way repeated-measures ANOVA, F(3.171, 193.4) = 51.12, P < 0.0001; trigger: F(4.544, 277.2) = 110.9, P < 0.0001), average speed (one-way repeated-measures ANOVA, F(3.969, 242.1) = 15.26, P < 0.0001). Bottom: from the left, entering latency (delay to enter the receptacle zone after the LED cue, one-way repeated-measures ANOVA, F(1.760, 107.4) = 9.652, P = 0.0003), zone occupancy (time spent in the receptacle zone after entering the zone during a trial, 3 s = maximum, one-way repeated-measures ANOVA, F(5.229, 277.1) = 3.420, P = 0.0045). Last three graphs depict success rate (one-way R repeated-measures ANOVA, F(1.727, 72.54) = 668.6, P < 0.0001), entering failure rate (one-way repeated-measures ANOVA, F(1.328, 55.79) = 244.5, P < 0.0001), and occupancy failure rate (one-way repeated-measures ANOVA, F(1.295, 54.40) = 61.04, P < 0.0001) comparisons for regular, reward-omission and rewarded LED-omission sessions of expert mice. n = 64 mice from all photometry behaviour experiments. Plotted as mean ± s.e.m. across mice and dots = mouse averages. ***P < 0.001 for Bonferroni-corrected post hoc comparisons. b, DAN activity across learning. The average responses for beginner, intermediate, expert, reward-omission (of expert mice) and rewarded LED-omission (of expert mice) trials are shown in red, orange, green, blue and purple, respectively. Dashed vertical lines indicate the behavioural time stamps (T = trigger zone entry, L = LED on, Z = receptacle zone entry, D = pellet dispensing, R = receptacle entry). Top: normalized Δf/f of VTA jRCaMP signal showing VTA DAN soma activity. Bottom: normalized Δf/f of NAc jRCaMP signal showing VTA DAN terminal activity. n = 10 mice. Plotted as mean ± s.e.m. across mice. c, VTA jRCaMP response (mean of normalized signal) to LED (mean of 0–3 s after LED cue) during training. Left, Individual mouse average plotted for different training periods (beginner, intermediate and expert) plotted as mean ± s.e.m. across trials (one-way repeated-measures ANOVA, F(1.870, 14.96) = 73.76, P < 0.0001). Right, daily average of LED response versus success rate, in which each dot represents a daily (session) measurement of a mouse plotted as linear regression fit ± its 95% confidence interval. d, VTA jRCaMP response to reward (mean of 3 s around the peak after receptacle entry) during training plotted as in c (one-way repeated-measures ANOVA, F(1.835, 14.68) = 40.40, P < 0.0001). e, VTA jRCaMP response in trained mice. Left, response to reward omission (RO-dip, mean of 3–6 s (shifted for slow soma jRCaMP signal) after expected time of pellet dispensing, one-sample t-test, P = 0.013) and to LED omission (LO-LED, mean of 0–3 s after expected time of LED onset, one-sample t-test, P = 0.045) plotted as mean ± s.e.m. across mice. Right, response to reward (mean of 0–3 s after pellet dispensing) in regular success (suc) and LED-omission (LO) trials (paired t-test, P = 0.001) plotted as in c. f, As in c, for NAc jRCaMP (one-way repeated-measures ANOVA, F(1.986, 15.89) = 98.52, P < 0.0001). g, As in d, for NAc jRCaMP (one-way repeated-measures ANOVA, F(1.939, 15.51) = 33.95, P < 0.0001). h, As in e, NAc jRCaMP response in trained mice. Left, response to reward omission (RO-dip, mean of 0–3 s after expected time of pellet dispensing, one-sample t-test, P < 0.0001) and to LED omission (LO-LED, mean of 0–3 s after expected time of LED onset, one-sample t-test, P = 0.189). Right, response to reward (mean of 0–3 s after pellet dispensing) in regular success (suc) and LED-omission (LO) trials (paired t-test, P < 0.0001). i, Daily average response of VTA jRCaMP, NAc jRCaMP and dLight response to LED and reward across training for individual mice. From the left, one-way mixed-effects analysis, F(1.589, 13.35) = 25.07, P < 0.0001; F(2.339, 18.71) = 11.70, P = 0.0003; F(1.720, 14.45) = 25.75, P < 0.0001; F(2.984, 23.87) = 15.69, P < 0.0001; F(1.806, 15.17) = 35.41, P < 0.0001; F(2.835, 22.68) = 12.73, P < 0.0001). Plotted as mean ± s.e.m.across trials for each mouse. *P < 0.05, **P < 0.01, ***P < 0.001 for one-sample t-tests, and paired t-tests, Bonferroni-corrected post hoc comparisons. All t-tests are two-sided.