Extended Data Fig. 9: Additional characterization of optogenetic manipulation of DA neural activity during defeat.

a, Arena schematic and example stimulation session trace from a 2-chamber real-time place preference assay. 20-Hz laser stimulation was delivered whenever a mouse’s centroid was in the “laser side” of the chamber, sides counterbalanced across mice (top). Preference for the laser-stimulation side of a chamber in mice expressing or not expressing excitatory opsin. No stimulation was delivered in the baseline condition (bottom). b, Animals receiving fight-back-triggered optogenetic activation expressed either ChRmine or ChR2 and were delivered 20 Hz laser stimulation with the following parameters: ChRmine mice received green laser stimulation in sets of 3 pulses without pause for the duration of fighting bouts while ChR2 mice received blue laser stimulation in sets of 10 pulses with a 0.5s pause between pulse sets for the duration of fighting bouts. c, Example of a defeat session with laser light delivery triggered on fighting of the closed-loop mouse and simultaneously delivered to his open-loop neighbor, bottom inset is a 10s segment. d, Statistics across fight-back-triggered stimulation: Distribution of latency to trigger stimulation from capture of a fighting video frame, with a median of 100ms (left). Distribution of durations of stimulation bouts, 94% of which were less than 1s (middle). Distribution of the amount of defeat time in which light was delivered (mean is 2.99% of the defeat session) (right). e, Attack-offset aligned laser stimulation during escape-triggered activation. f, In escape-triggered activation, distribution of the amount of defeat time in which light was delivered (mean is 0.75% of the defeat session). g, Time spent in random forest labeled fighting behavior early (days 1,2) versus late (days 9,10) in defeat, mean±SEM plotted. (Paired 2-sided t-Tests: 2-way FDR corrected: open-loop t = 0.38 p = 0.71, no opsin t = −0.41 p = 0.71; 2-way FDR corrected: escape-triggered t = −2.33 p = 0.10, no opsin t = −1.14 p = 0.34). h, Average expression of random forest labeled fleeing behavior early (days 1,2) versus late (days 9,10) in defeat, mean±SEM plotted. (Paired 2-sided t-Tests: 3-way FDR corrected: open-loop t = 1.12 p = 0.42, fight-triggered t = −0.29 p = 0.78, no opsin t = −1.49 p = 0.42). i, Percent of time immobile (speed < 1 cm/s) during exploration of a 2-chamber arena (2-sided t-Test, difference in mean immobility between open-loop (N = 14 males) and no opsin control (N = 14 males) group, mean±SEM plotted: t = −1.57, p = 0.12). j, Same as i for fight-back-triggered (N = 16 males) and no opsin (N = 14 males) controls, mean±SEM plotted (2-sided t-Test, t = −3.05, p = 0.005). k, Same as i for escape-triggered (N = 8 males) and control (N = 4 males) groups, mean±SEM plotted (2-sided t-Test, t = −0.80, p = 0.39). l, Schematic of inhibition of dopaminergic cell bodies in the VTA (Dio-NpHR injection and fiber implant over the VTA of DAT::Cre males). m, Real-time place aversion assay, with constant 7 mW green light delivery on the stimulation side of the chamber. Preference for the non-stimulation side of a chamber in mice expressing or not expressing inhibitory opsin, mean±SEM plotted. No stimulation was delivered in the baseline condition. n, Schematic for closed-loop, attack-triggered inhibition during defeat. o, In attack-triggered inhibition, distribution of the amount of defeat time in which light was delivered (mean is 7.93% of the defeat session). p, Density map of where in t-SNE behavior space animals were when receiving inhibition. q, Difference in occupancy of t-SNE clusters between inhibition (N = 12 males) and no opsin control (N = 10 males) groups (same mice in r–x), measured by individual clusters’ chi² statistics; black bold numbers label behaviors differentially expressed by resilient and susceptible mice in our observational study. r, Average time across 10 days of defeat being attacked (2-sided -Test, attack-triggered inhibition vs no opsin control, t = −3.03, p = 0.0066). s, Average time across 10 days of defeat fleeing (t = −3.92, p = 8.5E-4). t, Social interaction (SI) time in the SI test (2-sided t-Test, difference in SI time between inhibition and no opsin control groups: 2-sided t-Test, t = 0.04, p = 0.96). u, Average distance between stressed and aggressor mice during the first 5 min of the barrier-separated sensory period across the 10 days of defeat (2-sided Levene test for difference in variance, W = 4.92, p = 0.038). v, Number of entries into the open arms of an elevated plus maze (2-sided t-Test, difference in mean entries between inhibition and no opsin control, t = −0.64, p = 0.53). w, Number of crossings between chambers of 2-chamber arena (2-sided t-Test, difference in mean crossings between inhibition and no opsin control groups: t = −0.024, p = 0.98). x, Percent of time immobile (speed < 1 cm/s) during exploration of a 2-chamber arena (2-sided t-Test, difference in mean immobility between inhibition and no opsin control groups: t = −0.47, p = 0.64). In r–x, mean±SEM plotted. Across panels, *p < 0.05, **p < 0.01, ***p < 0.001.