Extended Data Fig. 1: Characterization of Sst⁺ CeA neuronal responses.

a, A schematic of the experimental setup and the approach. b, A raw image of the field of view under a GRIN lens. c, A confocal histological image of a coronal brain section from a representative mouse used for the imaging experiments, showing GCaMP6 expression in Sst⁺ CeA neurons and the location of GRIN lens implantation. d, Schematics showing the placement of GRIN lens implants in the mice (n = 16 mice) used for the imaging experiments. Note that some mice were used in more than one experiment, as indicated. e, Four example neurons showing selective responses to different stimuli. f, Heat-maps of the responses of all neurons to different stimuli. Each row represents the activities of one neuron. g, The first six principle components (PC) (explain 84% of the variance) and hierarchical clustering dendrogram showing the relationship of each neuron within the five clusters. h, Average responses of each cluster to different stimuli (n = 199 neurons/4 mice). i & j, Correlation coefficient matrixes of the responses of all neurons for each stimulus pair. k, The spatial locations of individual extracted neurons in the field of view (FOV) in the CeA of a representative mouse. Sst⁺ neurons excited by different stimuli are color coded. l, Cumulative probability distributions of the pairwise distances (measured as centroid distances) of neurons excited by different stimuli, as indicated, in the FOV. The distributions of the pairwise distances were significantly different between groups (water-excited neurons vs. sucrose-excited neurons, ****P = 1.33e-04; water-excited neurons vs. food-excited neurons, ****P = 1.06e-12; water-excited neurons vs. shock-excited neurons, ****P = 6.28e-13; sucrose-excited neurons vs. food-excited neurons, ****P = 1.39e-05; sucrose-excited neurons vs. shock-excited neurons, ****P = 2.40e-06; food-excited neurons vs. shock-excited neurons, n.s., nonsignificant, P = 0.34; Kolmogorov-Smirnov test). Data from 4 mice were pooled together (n = 243 cells/4 mice). m, Left: pie chart of the percentage distributions of Sst⁺ CeA neurons in naïve mice, showing those selectively excited by water or sound, or neither stimulus (nonresponsive). Right: the fractions of sound- or water-excited neurons in individual mice (n = 239 neurons/6 mice, *P = 0.0224, paired t-test). Sound intensity, 70 dB. n, Trial-by-trial activities of a representative water-excited neuron and the first 5 licking responses of the mouse in response to water delivery. Both the neural and the licking responses are aligned to the onset of water presentation (dashed line). o, The timing of increase in activity in the water-excited neuron shown in n, and the timing of increase in licking response of the corresponding mouse after the onset of water delivery, as determined by change-point analysis (Methods). Black dots represent the change-points. p, Quantification of the difference between neural and behavioral change-points, for all water-excited neurons and the corresponding mice. Values above zero indicate that neural responses lag behind behavioral responses. q & r, Histogram showing the distribution of water-excited neurons based on the trial-by-trial Pearson correlation coefficients between their activities and the licking rates of the mouse in the 3-s (q) or 10-s (r) time window immediately after the onset of water delivery (n = 10, r (8) = 0.64, P = 0.046; P > 0.05 when −0.64 < r < 0.64; t-test). s–x, Data acquired during the conditioning phase. s, Trial-by-trial activities of a representative CS_WA-excited neuron and the first 5 licking responses of the mouse in response to CS_WA. Both the neural and the licking responses are aligned to CS_WA onset. t, The timing of increase in activity in the neuron shown in s, and the timing of increase in licking response of the corresponding mouse after CS_WA onset, as determined by change-point analysis (Methods). Black dots represent the change-points. u, Quantification of the difference between neural and behavioral change-points, for all the responsive neurons and the corresponding mice. Values above zero indicate that neural responses lag behind behavioral responses. v, Histogram showing the distribution of neurons based on trial-by-trial Pearson correlation coefficients between their average activities and the licking rates of the mouse in the 2.5-s time window immediately after CS_WA onset. w, Histogram showing the distribution of neurons based on Pearson correlation coefficients between their US_WA responses and the CS_WA-induced licking rates of the mouse in subsequent trials (n = 19, r(17) = 0.46, P = 0.047; P > 0.05 when −0.46 < r < 0.46; t-test). x, Histogram showing the distribution of neurons based on Pearson correlation coefficients between their US_WA responses (in three-trial rolling averages) and the CS_WA-induced licking rates of the mouse (in the corresponding three-trial rolling averages) (n = 18, r(16) = 0.47, P = 0.049; P > 0.05 when −0.47 < r < 0.47; t-test). y-bb, Data acquired during the retrieval phase. y, Quantification of the difference between neural and behavioral change-points, for all the neurons excited by CS_WA and the corresponding mice. z, Histogram showing the distribution of neurons based on trial-by-trial Pearson correlation coefficients between their average activities and the licking rates of the mouse in the 3-s time window immediately after CS_WA onset (n = 20, r(18) = 0.45, P = 0.046; P > 0.05 when −0.45 < r < 0.45; t-test). aa, Quantification of the difference between neural and behavioral change-points, for all the neurons excited by CS_SH and the corresponding mice. bb, Histogram showing the distribution of neurons based on trial-by-trial Pearson correlation coefficients between their average activities and the eye size change of the mouse in the 3-s time window immediately after CS_SH onset (n = 15, r(13) = 0.52, P = 0.047; P > 0.05 when −0.52 < r < 0.52; t-test). cc-ff, data from the go/no-go task. cc, Quantification of the difference between neural and behavioral change-points, for all the neurons excited by CS_SU and the corresponding mice in hit trials. dd, Histogram showing the distribution of neurons based on trial-by-trial Pearson correlation coefficients between their average activities and the licking rates of the mouse in the 3-s time window immediately after CS_SU onset. ee, Quantification of the difference between neural and behavioral change-points, for all the neurons excited by CS_QU and the corresponding mice in false alarm (FA) trials. ff, Histogram showing the distribution of neurons based on trial-by-trial Pearson correlation coefficients between their average activities and the licking rates of the mouse in the 3-s time window immediately after CS_QU onset. Data are presented as mean ± s.e.m. Shaded areas represent s.e.m.

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