Conditional and unconditional components of aversively motivated freezing, flight and darting in mice
- Staglin Center for Brain and Behavioral Health, Department of Psychology, Department of Psychiatry and Biobehavioral Sciences, University of California, Los Angeles, United States
Figures
Figure 1 with 1 supplement
Behavioral design and schematic representation of the serial compound conditional stimulus (SCS) used for the Replication Group in Experiment 1.
During training, animals were given 2 days each of five SCS-US pairings. The SCS consisted of a 10 s pure tone (7.5 kHz) followed by a 10 s white noise (75 dB). Immediately upon termination of the white noise-SCS, a footshock US (1 s, 0.9 mA) was delivered. On day 3, the animals were tested with 16 presentations of the SCS without delivering any shocks.
Figure 1—figure supplement 1
Example traces of velocity (cm/s) measurements obtained via EthoVision across five trials on the first day of training for a mouse in the Replication Group of Experiment 1.
Vertical dotted lines denote stimulus onset times and the horizontal dotted line is the threshold for scoring behavior as a dart (22.9 cm/s). Darting episodes are marked with an *. See Figure 1—figure supplement 1—source data 1.
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Figure 1—figure supplement 1—source data 1
Source files for velocity used to create representative traces.
- https://cdn.elifesciences.org/articles/75663/elife-75663-fig1-figsupp1-data1-v2.xlsx
Figure 2 with 1 supplement
Trial-by-trial mean (±SEM) percent freezing, peak activity ratio, and darts per minute throughout all stimulus presentations during training (left panels) and testing (right panels) for the Replication Group (n=7; left half of figure) and the Stimulus Change Group (n=7; right half of figure) in Experiment 1.
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Figure 2—source data 1
Source files for freezing, PAR, and darting for Experiment 1-Replication and Stimulus Change groups.
- https://cdn.elifesciences.org/articles/75663/elife-75663-fig2-data1-v2.xlsx
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Figure 2—source data 2
Source files for velocity for all of Experiment 1.
- https://cdn.elifesciences.org/articles/75663/elife-75663-fig2-data2-v2.xlsx
Figure 2—figure supplement 1
Mean (±SEM) percent freezing, peak activity ratio, and darts per minute throughout training (left panels) and testing (right panels) for the CS Duration Group (n=8) of Experiment 1.
Responding during the 10 s preCS period is represented with open circles, responding during the noise is represented with filled in black circles. See Figure 2—figure supplement 1—source data 1.
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Figure 2—figure supplement 1—source data 1
Source files for freezing, PAR, and darting for Experiment 1-CS Duration group.
- https://cdn.elifesciences.org/articles/75663/elife-75663-fig2-figsupp1-data1-v2.xlsx
Figure 3
Averaged traces of velocity (cm/s) across the first four trials of extinction during testing for Experiment 1.
Data were averaged across all animals per group and binned into ~0.5 s bins (0.533 s) and presented as mean ± SE. These within-subject error bars are corrected for between-subject variability using methods as described in Morey, 2008. During this test, the Replication Group (n=7) and the Stimulus Change Group (n=7) received the serial conditional stimulus in which a 10 s tone was followed by a 10 s noise. The CS Duration Group (n=8) was only tested with a 10 s noise. See Figure 3—source data 1.
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Figure 3—source data 1
Source files for velocity for test day of Experiment 1.
- https://cdn.elifesciences.org/articles/75663/elife-75663-fig3-data1-v2.xlsx
Figure 4 with 1 supplement
Mean (± SEM) percent freezing, peak activity ratio, and darting for the test session for Experiment 2 (n=8 per group).
Values are averaged across the 16 trials of extinction during test. p-values and significance were determined through one-way ANOVA. *p<0.05, **p<0.01, ****p<0.0001. See Figure 4—source data 1 & Figure 4—source data 2.
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Figure 4—source data 1
Source files for freezing, PAR, and darting for test day for Experiment 2.
- https://cdn.elifesciences.org/articles/75663/elife-75663-fig4-data1-v2.xlsx
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Figure 4—source data 2
Source files for velocity for Experiment 2.
- https://cdn.elifesciences.org/articles/75663/elife-75663-fig4-data2-v2.xlsx
Figure 4—figure supplement 1
Trial-by-trial mean (± SEM) percent freezing, peak activity ratio, and darts per minute throughout 16 trials of testing for Experiment 2 (n=8 per group).
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Figure 4—figure supplement 1—source data 1
Source files for freezing, PAR, and darting for Experiment 2.
- https://cdn.elifesciences.org/articles/75663/elife-75663-fig4-figsupp1-data1-v2.xlsx
Figure 5
Averaged traces of velocity (cm/s) across the first four trials of extinction during testing for Experiment 2 (n=8 per group).
Data were averaged across all animals per group and binned into ~0.5 s bins (0.533 s) and presented as mean ± SE. These within-subject error bars are corrected for between-subject variability using methods as described in Morey, 2008. During this test, the no shock-noise test, shock only-noise test, and noise-shock noise test groups were tested with a 10 s noise. The shock only-tone test group was tested with a 10 s tone. See Figure 5—source data 1.
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Figure 5—source data 1
Source files for velocity for test day of Experiment 2.
- https://cdn.elifesciences.org/articles/75663/elife-75663-fig5-data1-v2.xlsx
Figure 6
Trial-by-trial mean (± SEM) percent freezing, peak activity ratio, and darting per minute throughout all stimulus presentations during training (left panels) and testing (right panels) for Experiment 3 (n=8 per group).
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Figure 6—source data 1
Source file for freezing, PAR, and darting for Experiment 3.
- https://cdn.elifesciences.org/articles/75663/elife-75663-fig6-data1-v2.xlsx
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Figure 6—source data 2
Source files for velocity for Experiment 3.
- https://cdn.elifesciences.org/articles/75663/elife-75663-fig6-data2-v2.xlsx
Figure 7
Averaged traces of velocity (cm/s) across two trials of extinction during testing for Experiment 3 (n=8 per group).
Datawere averaged across all animals per group and binned into ~0.5 s bins (0.533 s) and presented as Mean ± SE. These within-subject error bars are corrected for between-subject variability using methods as described in Morey, 2008. During this test all groups were tested with a 10 s noise CS.
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Figure 7—source data 1
Source files for velocity for test day of Experiment 3.
- https://cdn.elifesciences.org/articles/75663/elife-75663-fig7-data1-v2.xlsx
Figure 8 with 1 supplement
Averaged traces of velocity (cm/s) across three trials of extinction during testing for Experiment 4 (n=8 per group).
Datawere averaged across all animals per group and binned into ~0.5 s bins (0.533 s) and presented as mean ± SE. These within-subject error bars are corrected for between-subject variability using methods as described in Morey, 2008. During this test, all groups were tested with a 10 s noise CS. See Figure 8—source data 1.
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Figure 8—source data 1
Source files for velocity on test day of Experiment 4.
- https://cdn.elifesciences.org/articles/75663/elife-75663-fig8-data1-v2.xlsx
Figure 8—figure supplement 1
Trial-by-trial mean (± SEM) percent freezing, peak activity ratio, and darting per minute throughout all stimulus presentations during habituation (left panels), training (middle panels), and testing (right panels) for Experiment 4 (n=8 per group).
For habituation, presented are the values during CS presentation for habituation groups and values during the same 10 s period for context exposure groups. For training, presented are the values during CS exposure for the paired groups and values during the same 10 s period for shock only groups. For testing, all presented values are during the 10 s CS presentation. See Figure 8—figure supplement 1—source data 1 and Figure 8—figure supplement 1—source data 2.
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Figure 8—figure supplement 1—source data 1
Source files for freezing, PAR, and darting for all of Experiment 4.
- https://cdn.elifesciences.org/articles/75663/elife-75663-fig8-figsupp1-data1-v2.xlsx
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Figure 8—figure supplement 1—source data 2
Source files for velocity for all of Experiment 4.
- https://cdn.elifesciences.org/articles/75663/elife-75663-fig8-figsupp1-data2-v2.xlsx
Figure 9
Analysis of dart timing and magnitude.
(A) and (B) represent the magnitude of darts to the tone (n=48 darts) and noise (n=360 darts) stimuli during testing, as well as the reaction to the first shock (n=102 shocks) on day 1 of training. Data are presented as mean ± SE and come from all groups (total n=102 animals) that received shock during training, collapsed across all experiments. p-values and significance were determined through Welch’s ANOVA. *p<0.05, ****p<0.0001. (C) represents the magnitude (mean ± SE) of the first and second dart within a single CS presentation for all animals across all experiments that performed two darts within a single 10 s CS period (n=65 “multi-darts”). p-values and significance were determined through a paired sample t-test. *p<0.05. (D) and (E) represent the magnitude of darts that occurred during the initial 3 s of the 10 s CS period (n=230 darts) and those that occurred during the final 7 s of the 10 s CS period (n=178 darts). Data are presented as mean ± SE and come from all groups that received shock during training (n=102 animals), displayed by group and stimulus type in (D) and collapsed across all experimental groups in (E). p-values and significance were determined through Welch’s ANOVA. ****p<0.0001. See Figure 9—source data 1.
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Figure 9—source data 1
Source files for dart magnitudes and shock reactivity across all experiments.
- https://cdn.elifesciences.org/articles/75663/elife-75663-fig9-data1-v2.xlsx
Figure 10
Mean (± SEM) percent freezing during extinction/testing for Experiment 2 shows that the occurrence of the stimuli at test disrupt freezing to the context and that the noise disrupts freezing to a greater extent than the tone (n=8 per group).
Also plotted is a similar curve showing freezing and the impact that shock presentation during training has on freezing. These data are averaged across both shock only groups (total n=16) on day 2 trials after fear to the context had been established, showing that shock disrupts freezing to an even greater extent than the noise. See Figure 10—source data 1.
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Figure 10—source data 1
Source data for the ability of cues and shock to disrupt freezing.
- https://cdn.elifesciences.org/articles/75663/elife-75663-fig10-data1-v2.xlsx
Figure 11
Trial-by-trial mean (± SEM), peak activity ratio (PAR), and darting per minute throughout all stimulus presentations during training (left panels), and testing (middle panels) for all groups across experiments that received noise-shock pairings, grouped by sex of the animal (n=15 females; n=16 males).
The right panels show individual values for each animal’s average PAR and darts per minute across training and testing. p-values and significance were determined through repeated measures ANOVA, and the interaction was followed up with pairwise t-tests.
*p<0.05. See Figure 11—source data 1.
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Figure 11—source data 1
Source data for the potential sex differences in PAR and darting.
- https://cdn.elifesciences.org/articles/75663/elife-75663-fig11-data1-v2.xlsx
Tables
Table 1
Design of Experiment 1.
| Group | Training treatment: 10 CS-US pairings(5 per day) | Testing treatment(16 on 1 day) |
|---|---|---|
| (1) Replication | 10 s tone→10 s noise→1 s shock | 10 s tone→10 s noise |
| (2) CS duration | 10 s noise→shock | 10 s noise |
| (3) Stimulus change | 20 s tone→shock | 10 s tone→10 s noise |
Table 2
Design of Experiment 2.
| Group | 2-Day training treatment: | Testing treatment |
|---|---|---|
| (1) Pseudoconditioned noise-shock only noise test | 10 Shocks (1-mA, 1 s, 150–210 s intertrial interval) | 5 Noise presentations (10 s) |
| (2) Pseudoconditioned tone-shock only tone test | 10 Shocks (1-mA, 1 s, 150–210 s intertrial interval) | 5 Tone presentations (10 s) |
| (3) No shock control | Context exposure only (17 min and 15 s per day) | 5 Noise presentations (10 s) |
| (4) Noise-shock conditioning | 10 Noise (10 s)→shock pairings | 5 Noise presentations (10 s) |
Table 3
Design of Experiment 3—paired vs unpaired noise-shock.
| Group | Training treatment: 10 CS-US pairings(5 per day) | Testing treatment(2 on 1 day) |
|---|---|---|
| (1) Paired noise-shock (conditioning) | 10 s noise→1 s shock | 10 s noise |
| (2) Unpaired noise-shock | 10 s noise and 1 s shock – unpaired | 10 s noise |
| (3) Noise - CS only | 10 s noise | 10 s noise |
| (4) Shock only (pseudo conditioning) | 1 s shock | 10 s noise |
Table 4
Design of Experiment 4—tested the effect of habituation to the white noise.
| Group | Habituation treatment: 10 CS exposures (5 per day) | Training treatment: 10 CS-US pairings (5 per day) | Testing treatment(3 on 1 day) |
|---|---|---|---|
| (1) Habituation/shock only (H-shock) | 10 s noise | 1 s shock | 10 s noise |
| (2) Habituation/paired noise-shock (H-paired) | 10 s noise | 10 s noise→1 s shock | 10 s noise |
| (3) Context exposure/shock only (C-shock) | Context exposure | 1 s shock | 10 s noise |
| (4) Context exposure/paired noise-shock (C-paired) | Context exposure | 10 s noise→1 s shock | 10 s noise |
Additional files
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Transparent reporting form
- https://cdn.elifesciences.org/articles/75663/elife-75663-transrepform1-v2.docx
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Source code 1
Custom R code for darting analysis.
- https://cdn.elifesciences.org/articles/75663/elife-75663-code1-v2.zip
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Conditional and unconditional components of aversively motivated freezing, flight and darting in mice
eLife 11:e75663.
https://doi.org/10.7554/eLife.75663
