Behavioral generalization

Transcription

1 Supplementary Figure 1 Behavioral generalization. a. Behavioral generalization curves in four Individual sessions. Shown is the conditioned response (CR, mean ± SEM), as a function of absolute (main) or real (inset) distance from CS. The preparatory conditioned-response (CR) is the change in the inhale that triggers the CS tone, i.e. before the inhale that triggers odor release hence a preparatory response, and quantified as the half-height-time of the inhale in ms. It is then normalized within session to account for differences across days. b. The width of behavioral generalization (i.e. the half-height width from Fig.1e), but computed separately for the first third of recording sessions, the 2 nd, and the last 3 rd (mean±sem). This shows that the behavioral generalization was similar along the experimental period (p>0.1, ANOVA). c. Behavioral generalization, similar to main Fig.1e but here the CR is quantified as the peak of the preparatory inhale (mean ± SEM). d. Same as Fig.1g when the CR is quantified as the peak as in (c) (significant difference between aversive and pleasant response). e. Same as Fig.1h when the CR is quantified as the peak as in (c). The just-noticeable-difference (JND, 70.7% of the maximal response) computed here was 6.9%.

2 Supplementary Figure 2 Examples of tuning curves in the amygdala. a. Two examples of single-cells tuning curves. The middle panel shows the firing rate as a function of the nine presented tones in the generalization phase (mean ± SEM). Outer panels show raster plots overlaid by PSTH of individual responses to each tone. b. Three additional examples of single-cells tuning curves: firing rate as a function of the nine presented tones in the generalization phase (mean ± SEM, dots indicate FR in individual trials). Insets show the same generalization curves but as a function of absolute distance from the CS (mean ± SEM).

3 Supplementary Figure 3 Tuning-width changes with preferred-stimulus post-conditioning, but not pre-conditioning. a. Change in tuning width (width in post- minus pre-conditioning, mean±sem) as a function of PS distance from CS (p=0.03, F=2.96, ANOVA). This suggests that neurons generally had wide tuning pre-conditioning and narrowed/widened their tuning with direct relationship to the distance of their post-conditioning PS from the CS. b. The same analysis using PS from pre-conditioning showed no relationship (p=0.92 F=0.16 ANOVA). c. Tuning width during post-conditioning generalization as a function of the PS in the pre-conditioning habituation (phase/tone/interaction p=0.25/0.76/0.59 F=1.33/0.39/0.63, 2-way ANOVA). d. Same as in Fig.2e, but with width of tuning from pre-conditioning habituation (r=0.17, p=0.95, Pearson).

4 Supplementary Figure 4 There were no changes in tonic activity of BLA neurons. Firing rate during the pre-conditioning (habituation) plotted against the firing rate in the post-conditioning (generalization) phase for each neuron. Insets show the overall average in both phases (mean ± SEM). Left: activity taken from 1 sec before the tones and represents baseline activity. Right: activity taken from 1 sec after the tone and represents evoked response. There was no change in tonic responses (p=0.829 for left and for right, paired t-test).

5 Supplementary Figure 5 Additional characteristics of tuning in the post-learning phase. a. Correlation between Gaussian fit (R 2 ) and the absolute distance between the PS and the CS during generalization (r=-0.137, p=0.046; during habituation: r=-0.01, p=0.92). The slightly better fit for neurons with PS closer to the CS can reflect the narrower tuning described. b. Distribution of PS in pre- and post-conditioning periods. There was no change in variance of the distributions (right, variance ± CI, p>0.1), indicating that in the current paradigm, neurons did not shift their PS selectively towards the CS. c. Daily behavioral PS (center of Gaussian fit to the behavioral responses) varied with the neural PS recorded in the same session, but only during post-conditioning phase (left, r=0.38 p=0.005; pre-conditioning: right, r=-0.04 p=0.8).

6 Supplementary Figure 6 Neurons increase their tuning width rather than adding a CS response. a. Normalized firing rates for all tones as a function of absolute distance from the CS. Each panel depicts the average over all neurons with PS distance from the CS in a specific range (indicated in the color legend). All curves have a single peak at the PS, rather than a double peak at the PS and at the CS. b. Difference in (normalized) firing rate between the PS segment and the segment next to it, for the four groups of neurons as in (a). Inset shows the slope of these lines. The closer the PS to the CS, the steeper the slope (p<0.001, F=7.7, ANOVA). This strengthens the main finding, of narrower curves next to the CS.

7 Supplementary Figure 7 Basic auditory tuning properties of neurons in the primate amygdala. Pseudorandomized presentation of 40 pure-tones, ranging Hz (4 octaves) in equal logarithmic steps. Each tone was presented 30 times for 200ms (10ms rise/fall ramp; ITI:200ms). a. Rater plots overlaid with PSTH for 4 BLA neurons, aligned on tone onset. b. Auditory STRF for 4 BLA neurons (40 tones spanning 4 octaves in equal logarithmic distance) c. Distribution of number of peaks showing that most neurons had one best-frequency (BF, n=126). Zero-peaks was defined for broadly un-tuned units that responded equally to the entire tested spectrum. Inset shows the mean firing rate for the three different groups (mean ± SEM) d. BF distribution from tuned units was homogenous (logarithmic scale, χ 2 =25.7, p>0.1, n=99) e. Five examples of single-cells tuning. STRFs were averaged over the first 200 ms, and shown either in color scale or as a graph (upper three are 1-peak, fourth is 2-peak, lower is 0-peak). f. Distribution of response bandwidths and its cumulative distribution. Median=0.76 g. Latency distribution of response to each unit s PS. Median=76ms. Notice the relatively large group of neurons with latency of <50ms.

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9 Supplementary Figure 9 Robustness of relationship between width and PS distance from CS. a. Linear regression between tuning-width and PS distance from CS for neurons with different tuning fits. From left to right: for neurons with tuning-fit of R 2 > 0.1, >0.2, >0.3, >0.5, >0.6; R 2 >0.4 is shown in main Fig.2e. All regressions were significant with similar coefficients. b. The slope between the PS and the tuning-width, separately for the 1 st third of the recording epoch, the 2 nd, and the final third; showing similar neural changes following conditioning along the recording period.

10 Supplementary Figure 10 Behavioral and neural dynamics during conditioning and post-learning. a. Behavioral conditioned response (CR, the half-height-time of the inhale that triggers the CS, mean±sem) during the first third of the acquisition trials, the 2 nd, and the last third (F=5.16, p=0.002, ANOVA). b. Neural response (firing rate evoked by the CS, mean±sem) during the first third of the acquisition trials, the 2 nd, and the last third (F=2.76, p=0.04, ANOVA). c. Comparing rate of change for the neural response vs. the behavioral response on an individual basis (each dot is a neuron in one conditioning session). Upper: by fitting an exponential function to each and plotting λs; Lower: by fitting linear regression and plotting the slope. Solid lines represent the identity (upper: p=0.3, χ 2 =1.0; lower: p=0.04, χ 2 =4.5). d. Larger response for the CS that decays with distance from it, when counting spikes (left) or number of responding neurons (right). Insets show the same for the surrogate data.

11 Supplementary Figure 11 Tone discrimination based on neural activity. a. d between the CS and other tones, averaged over all neurons (top: experimental data; bottom: surrogate data), during habituation (grey) and generalization-test (black). This is the same figure as main Fig.3b inserted here for clarity. b. Change in d for all tone combinations. Color scale represents the difference in d from post-conditioning minus the d from preconditioning. The narrow diagonal band indicates no or negative improvement between near-by tones. c. To see how this result emerges from the population data, one can calculate the Fisher-Information (FI) for the CS when including neurons with PS farther from the CS and wide tuning curves (here, FI is the normalized average of the tuning-curves slope). This validated that information is high when considering only neurons with PS right next to the CS - because they have sharp tuning, hence their slope at the CS is large; but information decays when taking into account neurons with wide generalization curves because the information they provide is low due to a shallower slope around the CS.