Data from: Individual differences in fear memory expression engage distinct functional brain networks
Data files
Jun 30, 2026 version files 21.40 MB
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File_S01_Figure02A_statistics.csv
1.24 KB
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File_S02_Figure02B_statistics.csv
1 KB
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File_S03_Figure02_behavior_data.csv
193.26 KB
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File_S04_Figure06_PLS_brain_memory_behavior_wide.csv
116.06 KB
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File_S05_Figure06_boot_strap_ratios.csv
16.48 KB
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File_S06_Figure07_cfos_cell_counts_network_analysis.csv
194.18 KB
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File_S07_Figure08_network_confidence_intervals.csv
4.29 MB
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File_S08_Figure08_graph_evading_freezers.graphml
189.02 KB
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File_S09_Figure08_graph_freezers.graphml
198.74 KB
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File_S10_FigureS8-1_graph_non_reactive.graphml
102.29 KB
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File_S11_FigureS8-2_graph_vehicle.graphml
107.48 KB
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File_S12_Figure08_All_Brain_networks.cys
15.97 MB
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README.md
10.51 KB
Abstract
Fearful stimuli elicit a mix of active (e.g., evasion) and passive (e.g., freezing) behaviors in a wide range of species, including zebrafish (Danio rerio). However, it is not clear if individual differences in fear responses exist and what parts of the brain may underlie such differences. To probe these questions, we developed a contextual fear conditioning paradigm for zebrafish that uses conspecific alarm substance (CAS) as an unconditioned stimulus where animals associate CAS administration with a specific tank. To identify individual differences, we collected behavioral responses from over 300 fish from four different strains (AB, TU, TL, and WIK) and both sexes, finding that behavior fell into four distinct groups: non-reactive, evaders, evading freezers, and freezers. We also found that background strain and sex influenced how fish respond to CAS, with males more likely to increase evasive behaviors than females and the TU strain is more likely to be unresponsive during fear memory expression. Finally, we performed whole-brain activity mapping to identify the brain regions that are associated with different behavioral responses. All groups exposed to the tank (non-reactive, evading freezers, and freezers) had strong engagement of the telencephalon. However, regions beyond the telencephalon distinguished behavioral groups: animals that have high levels of freezing, but low levels of evasion, uniquely engage the cerebellum, preglomerular nuclei, and pretectal areas, whereas those fish that mix evasion with freezing engage the preoptic and hypothalamic areas. Taken together, these findings reveal that zebrafish exhibit individual differences in fear memory expression that are supported and the neural level by differential engagement of regions beyond the telencephalon.
Dataset DOI: 10.5061/dryad.sqv9s4nfh
Description of the data and file structure
Details for the methods of data generation are available in the associated manuscript.
Files and variables
File: File_S01_Figure02A_statistics.csv
Description: T-test statistics for data presented in Figure 2A
Variables
- Time.Strain.Sex: Time point (pre-exposure, CAS, or memory), strain, and sex of fish
- Freezing.p.value: p-values from t-tests comparing vehicle- and CAS-treated fish for percent freezing
- Freezing.q.value: q-values (FDR corrected p-values) from t-tests comparing vehicle and CAS treated fish for percent freezing
- Freezing.cohensD: Cohen's D comparing vehicle and CAS treated fish for percent freezing
- Evasion.p.value: p-values from t-tests comparing vehicle and CAS treated fish for percent evasive behavior as proportion of active time
- Evasion.q.value: q-values (FDR corrected p-values) from t-tests comparing vehicle and CAS treated fish for percent evasive behavior as proportion of active time
- Evasion.coshensD: Cohen's D from t-test comparing vehicle and CAS treated fish for percent evasive behavior as proportion of active time
File: File_S02_Figure02B_statistics.csv
Description: Statistics (one-sample t-tests) for Figure 2B
Variables
- Strain.Sex: Strain and sex of the fish
- p.value.freezing.CAS: p-values for percent freezing during CAS exposure relative to pre-exposure
- q.value.freezing.CAS: q-values (FDR corrected p-values) for percent freezing during CAS exposure relative to pre-exposure
- D.freezing.CAS: Cohen's D for percent freezing during CAS exposure relative to pre-exposure
- p.value.freezing.memory: p-values for percent freezing during memory day relative to pre-exposure
- q.value.freezing.memory: q-values (FDR corrected p-values) for percent freezing during memory day relative to pre-exposure
- D.freezing.memory: Cohen's D for percent freezing during memory day relative to pre-exposure
- p.value.per.evasive.CAS: p-values for percent of evasive swimming as proportion of non-freezing time during CAS exposure relative to pre-exposure
- q.value.per.evasive.CAS: q-values (FDR corrected p-values) for percent of evasive swimming as proportion of non-freezing time during CAS exposure relative to pre-exposure
- D.per.evasive.CAS: Cohen's D for percent of evasive swimming as proportion of non-freezing time during CAS exposure relative to pre-exposure
- p.value.per.evasive.memory: p-values for percent of evasive swimming as proportion of non-freezing time during memory relative to pre-exposure
- q.value.per.evasive.memory: q-values (FDR corrected p-values) for percent of evasive swimming as proportion of non-freezing time during memory relative to pre-exposure
- D.per.evasive.memory: Cohen's D for percent of evasive swimming as proportion of non-freezing time during memory relative to pre-exposure
File: File_S03_Figure02_behavior_data.csv
Description: Behavioral data for each fish across sex, strain, and timepoint.
Variables
- fish.id: internal fish ID number
- Timepoint: Timepoint in the task (i.e., pre-exposure, CAS exposure, or memory day)
- Strain: Background strain of fish
- Sex: Sex of fish
- DOF: Date of fertilization
- Line: Internal fish line number
- Position: Position in the tank
- Group: Experimental group (vehicle or CAS)
- Age(months): Age of fish in months
- Burst.Swimming: Percent time fish spent burst swimming
- Erratic.Movement: Percent time fish spent performing erratic movements
- Freezing: Percent time fish spent freezing
- Normal.Turn: Percent time fish spent doing normal turns
- Straight.Swimming: Percent time fish spent straight swimming
- Normal.swimming: Percent time fish spent doing normal turns or straight swimming
- Evasive.swimming: Percent time fish spent doing burst swimming or erratic movements
- Perc.active.normal: Percent time fish spent doing normal swimming when not freezing
- Per.active.evasive: Percent time fish spent doing evasive swimming when not freezing
File: File_S04_Figure06_PLS_brain_memory_behavior_wide.csv
Description: Data used for PLS analysis
Variables
- fish.id: Internal fish ID
- Group: Experimental group (vehicle or CAS treated)
- Sex: Sex of fish
- Community: Behavioral cluster for fish
- Normal.swimming: Percent time fish spent normal swimming (straight turns or normal turns)
- Freezing: Percent time fish spent freezing
- Evasive.swimming: Percent time fish spent in evasive swimming (burst swimming or erratic movements)
- Per.active.evasive: Percent time fish spent in evasive swimming as proportion of time not freezing
- ZL-MFN: normalized cfos cell densities for each brain region.
File: File_S05_Figure06_boot_strap_ratios.csv
Description: Bootstrap ratio results from PLS analysis
Variables
- Abbreviation: Brain region abbreviation
- V1: Bootstrap ratios for contrast 1
- V2: Bootstrap ratios for contrast 2
- V3: Bootstrap ratios for contrast 3
- V4: Bootstrap ratios for contrast 4
- Ont.2: Ontological level 2
- Ont.3: Ontological level 3
- Ont.4: Ontological level 4
- Display: Ontological level for display in figures
- IDX: Index number for region corresponding to the index used in the adult zebrafish brain atlas (AZBA)
File: File_S06_Figure07_cfos_cell_counts_network_analysis.csv
Description: cell count data used for the network analysis
Variables
- fish.id: internal fish ID
- Group: experimental group (vehicle or CAS)
- Sex: sex of fish
- cluster: Behavioral cluster of fish
- ZL-MFN: normalized cfos cell densities for each brain region.
File: File_S07_Figure08_network_confidence_intervals.csv
Description: correlations and confidence intervals used to determine the presence of overlapping or non-overlapping edges
Variables
- region.1: Abbreviation for brain region 1 used for generating the correlation
- region.2: Abbreviation for brain region 2 used for generating the correlation
- lower.Evading.freezer: lower bound of the 95% confidence interval for the correlation in the evading freezer network
- r.Evading.freezer: correlation in the evading freezer network
- upper.Evading.freezer: upper bound of the 95% confidence interval for the correlation in the evading freezer network
- p.Evading.freezer: p-value from the correlation in the evading freezer network
- lower.Freezer: lower bound of the 95% confidence interval for the correlation in the freezer network
- r.Freezer: correlation in the freezer network
- upper.Freezer: upper bound of the 95% confidence interval for the correlation in the freezer network
- p.Freezer: p-value from the correlation in the freezer network
- lower.Non.reactive: lower bound of the 95% confidence interval for the correlation in the non.reactive network
- r.None.reactive: correlation in the non-reactive network
- upper.Non.reactive: upper bound of the 95% confidence interval for the correlation in the none-reactive network
- p.Non.reactive: p-value from the correlation in the non-reactive network
- lower.Habituation: lower bound of the 95% confidence interval for the correlation in the habituation network
- r.Habituation: correlation in the habituation network
- upper.Habituation: upper bound of the 95% confidence interval for the correlation in the habituation network
- p.Habituation: p-value from the correlation in the habituation network
- fr.ef.sig.diff: whether the confidence intervals in the freezer and evading freezer networks overlap (1 = no overlap)
- fr.nr.sig.diff: whether the confidence intervals in the freezer and non-reactive networks overlap (1 = no overlap)
- ef.nr.sig.diff: whether the confidence intervals in the evading freezer and non-reactive networks overlap (1 = no overlap)
File: File_S08_Figure08_graph_evading_freezers.graphml
Description: Network file corresponding to the evading freezer group in Figure 8. Figure 8 depicts the functional network (derived from c-Fos-based neural activity measures) for the evading freezer and freezer groups. The nodes in the network correspond to brain regions, and the edges in the network correspond to the presence of suprathreshold (FDR-corrected p-value < 0.001) correlated activity between brain regions. Edges are also encoded based on whether the confidence interval for the correlated activity overlaps with the other two networks (freezers and non-reactive).
File: File_S09_Figure08_graph_freezers.graphml
Description: Network file corresponding to the freezer group in Figure 8. Figure 8 depicts the functional network (derived from c-Fos-based neural activity measures) for the evading freezer and freezer groups. The nodes in the network correspond to brain regions, and the edges in the network correspond to the presence of suprathreshold (FDR-corrected p-value < 0.001) correlated activity between brain regions. Edges are also encoded based on whether the confidence interval for the correlated activity overlaps with the other two networks (evading freezers and non-reactive).
File: File_S10_FigureS8-1_graph_non_reactive.graphml
Description: Network file corresponding to the non-reactive group in Figure S8-1. Figure S8-1 depicts the functional network (derived from cfos based neural activity measures) for the non-reactive group. The nodes in the network correspond to brain regions, and the edges in the network correspond to the presence of suprathreshold (FDR-corrected p-value < 0.001) correlated activity between brain regions. Edges are also encoded based on whether the confidence interval for the correlated activity overlaps with the other two networks (evading freezers and freezers).
File: File_S11_FigureS8-2_graph_vehicle.graphml
Description: Network file corresponding to the vehicle group in Figure S8-2. Figure S8-2B depicts the functional network (derived from cfos based neural activity measures) for the vehicle group. The nodes in the network correspond to brain regions and the edges in the network correspond to the presence of suprathreshold correlations (FDR-corrected p-value < 0.001) between brain regions.
File: File_S12_Figure08_All_Brain_networks.cys
Description: File for viewing networks in Cytoscape
Code/software
Visualization of graphml files is best done using Cytoscape
