Complementary stable and dynamic prelimbic ensembles encode learned threat value underlying generalization and discrimination
- Department of Psychological and Brain Sciences, University of Iowa, United States
- Psychology Department, University of Jaén, Campus Las Lagunillas, Spain
Figures
Figure 1 with 2 supplements
Experimental design, behavioral responses, viral strategy, and calcium imaging.
(a) Experimental design illustrating discriminative fear conditioning with either a 15 kHz CS+ or a 3 kHz CS+ (conditioned groups) and no-shock controls, which were tested at the same time points but never received footshocks. All groups were tested with 3 and 15 kHz tones and two intermediate frequencies (7 and 11 kHz) on days 1, 15, and 30 after conditioning. (b) Behavioral responses across retrieval sessions (CS+ 15 group: n=27 mice; CS+ 3 group: n=22 mice; no-shock controls: n=13 on days 1 and 15 and n=11 on day 30). Two control mice lost their lenses after day 15 and, therefore, did not contribute data to the day 30 session. The repeated-measures analysis included the 11 control mice with complete data across all sessions. Two-way repeated-measures ANOVA. CS+ 15 group: main effect of day, F(2,52) = 13.28, p<0.001; main effect of frequency, F(3,78) = 85.09, p<0.001; and day × frequency interaction, F(6,156) = 3.79, p=0.002. CS+ 3 group: main effect of day, F(2,42) = 14.42, p<0.001; main effect of frequency, F(3,63) = 58.81, p<0.001; and day ×frequency interaction, F(6,126) = 1.41, p=0.217. No-shock controls: main effect of day, F(2,20) = 1.38, p=0.276; main effect of frequency, F(3,30) = 1.43, p=0.253; and day × frequency interaction, F(6,60) = 0.335, p=0.916. Significant pairwise comparisons following Tukey’s multiple-comparisons test are denoted by asterisks: ***p<0.001. (c) Viral strategy and representative histological section showing GCaMP6f expression. Imaging depth did not exceed 300 μm, restricting recordings to the prelimbic cortex (PL). (d) Representative calcium-imaging data showing a raw fluorescence frame, processed frame, maximum-intensity projection across frames, and corresponding activity traces.
Figure 1—figure supplement 1
Discrimination ratios.
(a–c) Discrimination ratios (DRs) were used to quantify discrimination between each frequency and the CS+ across testing days in the CS+ 15 group (n=27) (a), CS+ 3 group (n=22) (b), and control animals (n=13) (c). In both experimental groups, discrimination increased over time, particularly between the CS+ and the CS− and between the CS+ and the frequency adjacent to the CS−. Discrimination between the CS+ and its adjacent frequency also increased across testing days in the CS+ 3 group but not in the CS+ 15 group (Tukey’s multiple comparisons). In contrast, control animals showed no consistent changes in discrimination over time. Repeated-measures ANOVAs with frequency comparison and testing day as factors revealed significant effects of frequency comparison, testing day, and their interaction in the CS+ 15 group (frequency: F(2,52) = 44.362, p<0.001; day: F(2,52) = 4.822, p=0.012; interaction: F(4,104) = 3.421, p=0.011). In the CS+ 3 group, significant effects of frequency comparison and testing day were observed, but not their interaction (frequency: F(2,42) = 24.415, p<0.001; day: F(2,42) = 5.499, p=0.008; interaction: F(4,84) = 0.23, p=0.921). No significant effects were detected in control animals (frequency: F(2,20) = 1.21, p=0.319; day: F(2,20) = 0.767, p=0.477; interaction: F(4,40) = 0.154, p=0.960). Asterisks denote Tukey’s multiple comparisons: p<0.05; **p<0.01; ***p<0.001.
Figure 1—figure supplement 2
Sex differences in behavior.
(a–c) Sex differences in CS+ 15-trained mice (n=27). No sex differences were observed on any testing day (effect of sex: day 1, F(1,25) = 1.52, p=0.23; day 15, F(1,25) = 0.55, p=0.467; day 30, F(1,25) = 0.17, p=0.681). On all days, there was a significant main effect of frequency, indicating successful learning in both males and females (day 1, F(3,75) = 33.46, p<0.001; day 15, F(3,75) = 26.32, p<0.001; day 30, F(3,75) = 51.47, p<0.001). No sex ×frequency interactions were detected on any testing day (p>0.05). Post hoc comparisons showed that on day 1, mice discriminated 3 kHz from 7, 11, and 15 kHz (p<0.05), but generalized among the higher frequencies (p>0.05). On days 15 and 30, mice discriminated 3 kHz from 7, 11, and 15 kHz (p<0.05) and 7 kHz from 11 and 15 kHz (p<0.05), while generalization persisted between 11 and 15 kHz (p>0.05). (d-f) Sex differences in CS+ 3-trained mice (n=22). No main effect of sex was observed on any testing day (day 1, F(1,20) = 1.24, p=0.279; day 15, F(1,20) = 0.78, p=0.388; day 30, F(1,20) = 0.003, p=0.956). A significant main effect of frequency was present across all days, indicating learning in both sexes (day 1, F(3,60) = 40.65, p<0.001; day 15, F(3,60) = 30.17, p<0.001; day 30, F(3,60) = 35.19, p<0.001). Post hoc comparisons indicated discrimination among all frequencies across days (p<0.05), except between 11 and 15 kHz (p>0.05). Significant sex ×frequency interactions were observed on day 1 (F(3,60) = 6.91, p<0.001) and day 15 (F(3,60) = 3.55, p<0.02), reflecting sex-specific differences at 11 kHz on day 1 and at 3 kHz on day 15. These interaction effects were not present on day 30 (F(3,60) = 0.76, p=0.522) and likely reflect increased behavioral variability rather than stable sex differences.
Figure 2 with 1 supplement
Distribution of stable and dynamic cell types across sessions.
(a) Activity plots from a representative animal conditioned with a 15 kHz CS+, with neurons ordered by activity level. The plots show positive tone-responsive neurons (bottom), negative tone-responsive neurons (top), and mixed or nonresponsive neurons (middle) before and after tone presentation. (b) Proportions of neuronal response types across experimental groups and sessions. Numbers of cells–CS+ 15 group: conditioning, 3834; day 1, 3177; day 15, 4186; and day 30, 3693. CS+3 group: conditioning, 1628; day 1, 1381; day 15, 1881; and day 30, 1833. No-shock controls: conditioning, 1,118; day 1, 986; day 15, 1386; and day 30, 563. Two control mice yielded data only through day 15. (c) Venn diagrams showing the proportions of consistently active neurons (active during all three retrieval sessions), partially active neurons (active during two sessions), and transiently active neurons (active during only one session). The proportions of neurons in these categories were similar across groups, except that transient neurons on day 30 were more abundant in the control group. This difference reached significance only between the control and CS+ 15 groups (p<0.05; Supplementary file 1). Diagrams and analyses for the control group included only mice recorded through day 30.
Figure 2—figure supplement 1
Cell registration quality.
(a) Distribution of equivalent neuronal footprint diameters (µm) for all registered neurons. (b) Representative spatial footprints recorded on day 1 (red), day 15 (green), and day 30 (blue), along with the merged image showing neurons tracked throughout the experiment (white). (c) Total number of neurons recorded in each experimental group and the percentage of neurons successfully registered across all sessions. Note that two control animals were excluded because they lost the GRIN lens after day 15. (d–f) Distributions of centroid shifts between matched neurons in the retrieval sessions relative to the conditioning session for the CS+ 3 (d), CS+ 15 (e), and control (f) groups. The red line indicates the median equivalent neuronal footprint diameter (21.4 µm). The upper left corner of each panel shows the median centroid shift and the percentage of registered neurons with centroid shifts smaller than the median footprint diameter.
Figure 3 with 3 supplements
Population activity of positive tone-responsive neurons exhibits graded patterns reflecting learned threat value.
(a–c) Population responses in animals conditioned with a 15 kHz CS+ (n=7) (a), animals conditioned with a 3 kHz CS+ (n=5) (b), and no-shock controls (n=6) (c). In each group, the upper panels show positive tone-responsive neurons, and the lower panels show negative tone-responsive neurons. Adjacent boxplots show the areas under the population response curves (AUCs). Boxplots indicate the median (center line) and interquartile range (box); whiskers extend to the most extreme values within 1.5 times the interquartile range, and points beyond the whiskers represent individual outliers. CS+ 15 group–positive responders: day 1, F(3,18) = 9.963, p<0.001; day 15, F(3,18) = 9.973, p<0.001; and day 30, F(3,18) = 6.627, p=0.003. Negative responders: day 1, F(3,18) = 2.483, p=0.094; day 15, F(3,18) = 1.877, p=0.178; and day 30, F(3,18) = 2.753, p=0.073. CS +3 group–positive responders: day 1, F(3,12) = 6.899, p=0.006; day 15, F(3,12) = 9.247, p=0.002; and day 30, F(3,12) = 6.123, p=0.009. Negative responders: day 1, F(3,12) = 0.870, p=0.484; day 15, F(3,12) = 0.512, p=0.641; and day 30, F(3,12) = 1.448, p=0.278. No-shock controls–positive responders: day 1, F(3,15) = 0.527, p=0.670; day 15, F(3,15) = 1.852, p=0.181; and day 30, F(3,9) = 1.046, p=0.418. Negative responders: day 1, F(3,13) = 1.205, p=0.347; day 15, F(3,13) = 1.375, p=0.294; and day 30, F(3,9) = 0.950, p=0.457. Asterisks denote significant Tukey post hoc comparisons: *p<0.05, **p<0.01, and ***p<0.001.
Figure 3—figure supplement 1
Population activity of consistently active neurons.
(a–b) Population activity of consistently active neurons from animals trained with a 15 kHz CS+ (n=7) (a) or a 3 kHz CS+ (n=5) (b). Upper panels show positively tone-responsive neurons and lower panels show negatively tone-responsive neurons. Adjacent boxplots display areas under the population response curves (AUC). Boxplots show the median (center line), interquartile range (box), and whiskers extending to ±1.5 × the interquartile range; points outside the whiskers represent individual observations beyond this range. Consistently active positive responders exhibit graded responses across test days (day 1 to day 30). *p<0.05; **p<0.01.
Figure 3—figure supplement 2
Population activity of emerging-retained neurons.
Emerging-retained neurons become part of the active ensemble after conditioning and remaining active after that. (a–b) Population activity of emerging-retained neurons from animals trained with a 15 kHz CS+ (n=7) (a) or a 3 kHz CS+ (n=5) (b). Upper panels show positively tone-responsive neurons and lower panels show negatively tone-responsive neurons. Adjacent boxplots display areas under the population response curves. Emerging-retained neurons show greater variability on day 1 compared with later test days. Only positive responders show significant graded valence. *p<0.05; **p<0.01.
Figure 3—figure supplement 3
Population activity of transiently active neurons.
Transiently active neurons were active on only a single test day. (a–b) Population activity of transiently active neurons from animals trained with a 15 kHz CS+ (n=7) (a) or a 3 kHz CS+ (n=5) (b). Upper panels show positively tone-responsive neurons and lower panels show negatively tone-responsive neurons. Adjacent boxplots display areas under the population response curves. Transiently active neurons did not show graded population responses on day 1; graded responses emerged by day 15 and were maintained through day 30 in positive sound responder neurons. *p<0.05; **p<0.01.
Figure 4 with 1 supplement
Generalized linear model (GLM) analysis of the relative contributions of tone identity and freezing behavior to neuronal activity for positive (a–d) and negative (e–h) responder cells.
(a) Regression coefficients (β) for positive tone-selective neurons in conditioned mice exhibit opposite monotonic gradients across tones after controlling for freezing. This graded pattern is absent in no-shock controls. (b) The freezing-to-tone coefficient ratio remains below 1 for every tone in all groups, indicating a greater contribution of tone identity than freezing to the activity of positive tone-selective neurons. (c) Within-cell comparisons confirm that the absolute median β coefficients for tone exceed the absolute freezing coefficients in all groups. (d) When pooled across tones, the proportion of freezing-dominant neurons remains stable across retrieval sessions. (e) Negative tone-selective neurons exhibit suppressive tone coefficients across all frequencies, with only weak frequency dependence, indicating the absence of pronounced graded responses. (f) Freezing-to-tone coefficient ratios are higher than those observed in positive responders but remain below 1 across tones in conditioned mice. In no-shock controls, the ratio approaches 1, indicating similar contributions of freezing and tone identity. (g) Within-cell comparisons confirm that the absolute tone coefficients exceed the absolute freezing coefficients, but only in the conditioned groups. (h) The proportion of freezing-dominant neurons remains relatively stable across retrieval sessions in each group. Controls displayed a slightly higher proportion of freezing-dominant neurons, but this difference was not significant. Significant Tukey multiple comparisons are denoted by asterisks: *p<0.05, **p<0.01, and ***p<0.001. CS+ 15: n=7; CS+ 3: n=5; No Shock: n=6.
Figure 4—figure supplement 1
Proportion of freezing-dominant neurons.
(a–b) Percentage of freezing-dominant neurons responding to each tone among positive (a) and negative (b) responders. A repeated-measures ANOVA, with group as the between-subjects factor and frequency as the within-subjects factor, revealed a significant group ×frequency interaction [F(6, 45)=2.402, p<0.05]. No main effects of group or frequency were observed (p>0.05). However, Tukey-corrected multiple comparisons revealed no significant pairwise differences, with only trends at 3 kHz between the CS+ 15 and CS+ 3 groups (p=0.087) and at 11 kHz between the CS+ 15 and no-shock groups (p=0.069). CS+ 15: n=7; CS+ 3: n=5; No Shock: n=6.
Figure 5
Population-vector similarity across tones and time.
(a–c) Population-similarity maps for all tone pairs across the time course of tone presentation in the CS+ 15 (a), CS+ 3 (b), and no-shock control (c) groups. (d) Schematic illustrating the color scale and orientation of the similarity maps. The y-axis represents the earlier tone in each comparison, and the x-axis represents the later tone. (e, f) Boxplots showing population similarity during the first 5 s after tone onset, quantified relative to the CS+, for the CS+ 15 (n=7) (e; F(3,36) = 12.025, p<0.001) and CS+ 3 (n=5) (f); F(3,24) = 7.435, p=0.004 groups. Asterisks denote significant Tukey post hoc comparisons: *p<0.05, **p<0.01, and ***p<0.001.
Figure 6 with 4 supplements
Signed mutual information (MI) clustering identifies prelimbic cortex (PL) subpopulations with distinct tone-response profiles.
(a) MI-based clustering pipeline. An MI matrix generated from simultaneously recorded PL neurons was spectrally clustered according to shared information, independently of response sign. Within each primary cluster, MI values were combined with the signs of pairwise correlations and reclustered to identify subclusters with distinct positive or negative activity patterns. Subcluster labels were then combined to generate the final signed MI matrix. (b) Clustered signed MI matrices for the CS+ 15 (top), CS+ 3 (middle), and no-shock control (bottom) groups, with neurons sorted by cluster label. Color indicates the sign and magnitude of MI. (c) Mean stimulus-aligned responses of clusters positively modulated by individual tones. (c.1–c.4) correspond to 3, 7, 11, and 15 kHz, respectively. (d) Mean stimulus-aligned responses of clusters negatively modulated by individual tones. (d.1–d.4) correspond to 3, 7, 11, and 15 kHz, respectively.
Figure 6—figure supplement 1
Prelimbic cortex (PL) subnetwork clusters identified using signed mutual information for the CS+ 15 group in each testing session.
(a–b) Average stimulus-aligned population responses for clusters showing positive (a) or negative (b) modulation to individual tones (3, 7, 11, or 15 kHz, upper panels) or graded emotional tuning (a-b, bottom panels).
Figure 6—figure supplement 2
Prelimbic cortex (PL) subnetwork clusters identified using signed mutual information for the CS+ 3 group in each testing session.
(a–b) Average stimulus-aligned population responses for clusters showing positive (a) or negative (b) modulation to individual tones (3, 7, 11, or 15 kHz, upper panels) or graded emotional tuning (a-b, bottom panels).
Figure 6—figure supplement 3
Prelimbic cortex (PL) subnetwork clusters identified using signed mutual information for the no-shock control group in each testing session.
(a–b) Average stimulus-aligned population responses for clusters showing positive (a) or negative (b) modulation to individual tones (3, 7, 11, or 15 kHz).
Figure 6—figure supplement 4
Baseline-to-stimulus firing rate ratio (BSR) changes in positively responding neurons within tone-specific clusters for the CS+ 15 (red) (n=7), CS+3 (blue) (n=5), and control groups (n=6).
(a) BSR of neurons in clusters primarily responsive to 3 kHz (Figure 5 c.1), (b) 7 kHz (Figure 5 c.2), (c) 11 kHz (Figure 5 c.3), and (d) 15 kHz (Figure 5 c.4). ANOVA results are reported in Supplementary file 3 Tukey’s multiple-comparison tests indicate significance (p<0.05; p<0.01; p<0.001).
Figure 7
Graded tone-responsive neurons are present only in conditioned groups.
(a, b) Mean stimulus-aligned population responses of clusters exhibiting graded tone responses in animals conditioned with a 15 kHz CS+, showing positive (a) and negative (b) response patterns. (c–e) Mean stimulus-aligned population responses of clusters exhibiting graded tone responses in animals conditioned with a 3 kHz CS+, showing positive (c, d) and negative (e) response patterns. (f, g) Stability of neuronal identity within graded tone-responsive clusters across retrieval sessions. Asterisks denote significance after Benjamini–Hochberg correction. (h, i) Baseline-to-stimulus firing-rate ratios (BSRs) showing changes in the activity of graded clusters across retrieval sessions in the CS+ 15 (n=7) (h) and CS+ 3 (n=5) (i) groups. Graded clusters were not identified in no-shock controls. Asterisks denote significant Tukey post hoc comparisons: *p<0.05, **p<0.01, and ***p<0.001.
Figure 8
Generalized linear model (GLM) analysis of the contributions of tone identity and freezing to the activity of clustered prelimbic cortex (PL) neurons.
(a–h) Positive (a–d) and negative (e–h) single-tone-selective neurons. (a) Tone β coefficients in conditioned mice form opposing gradients that peak at the CS+; these gradients are absent in no-shock controls. (b) Freezing-to-tone ratios remain below 1 across preferred tones and retrieval sessions. (c) Within-cell comparisons confirm that absolute tone coefficients exceed absolute freezing coefficients in all groups. (d) The proportion of freezing-dominant neurons is low and stable across retrieval sessions. (e) Negative tone-selective neurons exhibit suppressive tone β coefficients with weak frequency dependence and greater variability in no-shock controls. (f) Freezing-to-tone ratios are higher than those of positive responders but remain below 1 in conditioned mice. (g) Within-cell comparisons show that absolute tone coefficients exceed absolute freezing coefficients in negative responders from conditioned mice, although by a smaller margin than in positive responders. (h) The proportion of freezing-dominant neurons is higher than among positive responders but remains stable across retrieval sessions. (i–p) Positive (i–l) and negative (m–p) graded neurons. (i) Tone β coefficients obtained after controlling for freezing exhibit strong CS+-centered gradients. (j) Freezing-to-tone ratios are well below 1, indicating that tone identity contributes substantially more strongly to neuronal activity than freezing. (k) Within-cell comparisons confirm that absolute tone coefficients greatly exceed absolute freezing coefficients. (l) Only a small and stable proportion of positive graded neurons is freezing-dominant. (m) Negative graded neurons exhibit strong suppressive gradients. (n) Freezing-to-tone ratios remain below 1. (o) Absolute tone coefficients are approximately threefold greater than absolute freezing coefficients. (p) The proportion of freezing-dominant neurons remains low across retrieval sessions. Asterisks denote significant Tukey post hoc comparisons: *p<0.05, **p<0.01, and ***p<0.001. CS+ 15: n=7; CS+ 3: n=5; No Shock: n=6.
Tables
Key resources table
| Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Strain, strain background (Mus musculus, female and male) | C57BL/6 J | The Jackson Laboratory | RRID:IMSR_JAX_000664 | Age: 8–10 weeks. |
| Recombinant DNA reagent | AAV1-CaMKIIα-GCaMP6f-WPRE-SV40 | Addgene | RRID:Addgene_100834 | Calcium indicator delivered to the prelimbic cortex; three 150 nL injections; viral titer: 2×10¹³ GC/mL. |
| Chemical compound, drug | Isoflurane, USP | Piramal Critical Care, Inc. | NDC:66794-013-25 | 99.9% isoflurane; 250 mL bottle. Used at 3% for induction and 1.5% for maintenance during stereotaxic surgery. |
| Chemical compound, drug | Rimadyl (carprofen) | Zoetis Inc. | UPC:087219064702 | Injectable solution, 50 mg/mL; 20 mL vial. Administered at 5 mg/kg, s.c., preoperatively and on two consecutive postoperative days. |
| Software, algorithm | FreezeFrame 5 | Actimetrics | RRID:SCR_014429 | Freezing quantification in non-implanted mice. Version 5. |
| Software, algorithm | Inscopix Data Processing Software (IDPS) | Inscopix | Version: 1.9.8 | Preprocessing, motion correction and CNMF-E extraction of neuronal footprints and calcium traces. |
| Software, algorithm | CASCADE | Rupprecht et al., 2021 | https://doi.org/10.1038/s41593-021-00895-5 | Spike inference from calcium signals for population similarity and baseline-to-stimulus ratios. |
| Software, algorithm | Custom analysis code | This paper | https://doi.org/10.17632/9yyn63g346.3 | Sound-response classification, stimulus-aligned analyses, mutual-information clustering, cluster stability, IMU-based freezing estimates and GLM analyses. |
| Other | nVoke 2.0 miniscope system | Inscopix | RRID:SCR_023028 | Longitudinal calcium imaging and integrated inertial measurement unit recordings. |
| Other | GRIN lens | Inscopix | Model:1050–004637 | Diameter: 1.0 mm; length: 4.0 mm; numerical aperture: 0.5. |
Additional files
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Supplementary file 1
Statistical analyses corresponding to the Venn diagrams shown in Figure 2.
One-way ANOVAs were used when the assumptions of normality and homogeneity of variance were met; otherwise, Kruskal–Wallis tests were performed. Significant Kruskal–Wallis tests were followed by Dunn’s multiple-comparisons tests. Effect sizes are reported as partial eta squared (ηp²) for ANOVAs and epsilon squared (ε²) for Kruskal–Wallis tests.
- https://cdn.elifesciences.org/articles/111177/elife-111177-supp1-v1.docx
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Supplementary file 2
Statistics comparing areas under the curve for the CS+ 15, CS+ 3, and No shock control groups across all frequencies and testing sessions.
(a-b) Three-way mixed-model ANOVAs using Group (CS+ 15, CS+ 3, No shock), Frequency (3, 7, 11, 15 kHz), and Time (days 1, 15, 30) as variables for positive (a) and negative (b) cell responders. Simple effects were calculated using Holm correction.
- https://cdn.elifesciences.org/articles/111177/elife-111177-supp2-v1.docx
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Supplementary file 3
Statistics corresponding to consistently active, emerging-retained, and transiently active cells.
One-way ANOVAs with repeated measures were used when normality and equal variance tests were passed to test the effect of frequency. Friedman tests were used (χ² with Kendall’s W) when assumptions were violated, Friedman tests were used (χ² with Kendall’s W).
- https://cdn.elifesciences.org/articles/111177/elife-111177-supp3-v1.docx
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Supplementary file 4
Statistical analysis of generalized linear model (GLM)-derived measures for cells used in the population curves shown in Figure 4.
Two-way ANOVAs with repeated measures were conducted to evaluate effects of group (CS+ 15, CS+ 3, or No shock), frequency (3, 7, 11, 15 kHz), or day (days 1, 15, 30), and their interactions. Tukey post hoc comparisons are shown where simple effects were tested.
- https://cdn.elifesciences.org/articles/111177/elife-111177-supp4-v1.docx
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Supplementary file 5
Statistical analyses identifying neuronal clusters that remained stable over time, defined as neurons that retained the same functional response profile on days 15 and 30 as on day 1.
These analyses correspond to the clusters shown in Figures 6 and 7, as indicated in the table. For each cluster, the observed proportion of neurons that preserved their response profile was compared with a null distribution, with multiple comparisons controlled using the Benjamini–Hochberg procedure. Only the graded neuronal clusters in each experimental group (Figure 7) exhibited significant stability across time. No graded clusters were identified in the no-shock control group.
- https://cdn.elifesciences.org/articles/111177/elife-111177-supp5-v1.docx
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Supplementary file 6
Statistical analyses corresponding to the normalized baseline-to-stimulus firing rate (BSR) for the neuronal clusters shown in Figure 4—figure supplement 1, Figure 6—figure supplement 2.
BSRs for the clusters shown in Figure 4—figure supplement 1, Figure 6—figure supplement 2 are presented in Figure 6—figure supplement 3, whereas BSRs for the graded clusters shown in Figure 4—figure supplement 1, Figure 6—figure supplement 1 are presented in Figure 7h–i. For pure tone-selective clusters, two-way repeated-measures ANOVAs were used to evaluate the effects of group (CS+ 15, CS+ 3, or No Shock), frequency (3, 7, 11, and 15 kHz), and their interaction. For graded clusters, one-way repeated-measures ANOVAs were used to evaluate the effect of time. Tukey’s multiple-comparisons tests are reported in Figure 6—figure supplement 1 and Figure 7h–i.
- https://cdn.elifesciences.org/articles/111177/elife-111177-supp6-v1.docx
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Supplementary file 7
Statistical analysis of positive and negative modulated single-frequency and graded neurons shown in Figure 8.
Two-way ANOVAs with repeated measures were conducted to evaluate effects of group (CS+ 15, CS+ 3, or No shock), frequency (3, 7, 11, 15 kHz), day, condition (tone vs. freezing), and their interactions. Tukey post hoc comparisons are shown where simple effects were tested.
- https://cdn.elifesciences.org/articles/111177/elife-111177-supp7-v1.docx
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MDAR checklist
- https://cdn.elifesciences.org/articles/111177/elife-111177-mdarchecklist1-v1.pdf
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Complementary stable and dynamic prelimbic ensembles encode learned threat value underlying generalization and discrimination
eLife 15:RP111177.
https://doi.org/10.7554/eLife.111177.4
