Computational modeling reveals cognitive processes in simple rodent depression tests methods
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mouse
Subject model for the experiment.
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- confirm full cohort details in the source paper
Chemogenetic inhibition of medial prefrontal cortex (mPFC) pyramidal neurons
reagent used in the protocol.
- Use
- Stereotaxic surgeries were performed under general anesthesia with 1% sodium pentobarbital using a stereotaxic apparatus (RWD Life Science Co., Ltd, Shenzhen, China). To inhibit pyramidal neurons in the mPFC, 500nL of rAAV-CaMKIIα-hM4D(Gi)-mCherry (BrainVTA Co., Ltd, Wuhan, China) was bilaterally injected into...
Histology and microscopy
reagent used in the protocol.
- Use
- Following the final day of the behavioral experiments, mice were injected with 0.2 mL of a urethane solution at a concentration of 0.3 mg/mL to achieve anesthesia. Subsequently, they underwent transcardial perfusion with 4% paraformaldehyde in PBS for fixation. The brains were then post-fixed overnight, and coronal...
Chemogenetic inhibition of medial prefrontal cortex (mPFC) pyramidal neurons
Stereotaxic surgeries were performed under general anesthesia with 1% sodium pentobarbital using a stereotaxic apparatus (RWD Life Science Co., Ltd, Shenzhen, China). To inhibit pyramidal neurons in the mPFC, 500nL of rAAV-CaMKIIα-hM4D(Gi)-mCherry (BrainVTA Co., Ltd, Wuhan, China) was bilaterally injected into...
- Use
- Stereotaxic surgeries were performed under general anesthesia with 1% sodium pentobarbital using a stereotaxic apparatus (RWD Life Science Co., Ltd, Shenzhen, China). To inhibit pyramidal neurons in the mPFC, 500nL of rAAV-CaMKIIα-hM4D(Gi)-mCherry (BrainVTA Co., Ltd, Wuhan, China) was bilaterally injected into...
Behavioral tasks
The FST was performed in a well-lit room. Mice were individually placed in water-filled cylinders (35 cm in height × 10 cm in diameter) with water maintained at 24°C-25°C and at depth sufficient to prevent their limbs from touching the bottom. Each mouse was allowed to swim freely for 6 min, and...
- Use
- The FST was performed in a well-lit room. Mice were individually placed in water-filled cylinders (35 cm in height × 10 cm in diameter) with water maintained at 24°C-25°C and at depth sufficient to prevent their limbs from touching the bottom. Each mouse was allowed to swim freely for 6 min, and...
Behavioral tasks
For the TST, mice were gently removed from their cages, and their tails were secured with adhesive tape approximately 1 cm from the tip. The mice were then suspended by their tails at a height of about 30 cm from the ground, placing them in an inverted position. The test also lasted for 6 min, and video recordings w...
- Use
- For the TST, mice were gently removed from their cages, and their tails were secured with adhesive tape approximately 1 cm from the tip. The mice were then suspended by their tails at a height of about 30 cm from the ground, placing them in an inverted position. The test also lasted for 6 min, and video recordings w...
Histology and microscopy
Following the final day of the behavioral experiments, mice were injected with 0.2 mL of a urethane solution at a concentration of 0.3 mg/mL to achieve anesthesia. Subsequently, they underwent transcardial perfusion with 4% paraformaldehyde in PBS for fixation. The brains were then post-fixed overnight, and coronal...
- Use
- Following the final day of the behavioral experiments, mice were injected with 0.2 mL of a urethane solution at a concentration of 0.3 mg/mL to achieve anesthesia. Subsequently, they underwent transcardial perfusion with 4% paraformaldehyde in PBS for fixation. The brains were then post-fixed overnight, and coronal...
The swim struggle tracker (SST)
In brief, the SST consists of four modules: the calibration and region of interest (ROI) selection module, the passive motion compensation module, the active motion detection module, and the smooth and thresholding analysis module. We briefly describe the function of each module below. (1) Calibration and ROI select...
- Use
- In brief, the SST consists of four modules: the calibration and region of interest (ROI) selection module, the passive motion compensation module, the active motion detection module, and the smooth and thresholding analysis module. We briefly describe the function of each module below. (1) Calibration and ROI select...
The swim struggle tracker (SST)
Active motion detection module: As shown in B, active movement by the animal results in significant differences between adjacent frames, even after passive motion compensation. We use a three-frame differencing method to calculate inter-frame differences based on the ROI images from frames t and t+1. These differenc...
- Use
- Active motion detection module: As shown in B, active movement by the animal results in significant differences between adjacent frames, even after passive motion compensation. We use a three-frame differencing method to calculate inter-frame differences based on the ROI images from frames t and t+1. These differenc...
The swim struggle tracker (SST)
Behavioral trajectory smoothing and binning module: As illustrated in C, the original behavioral trajectory for each frame often contains significant noise. For example, in a video where a mouse struggles for the first 6 s and remains immobile for the following 6 s, initial recognition may produce noisy results, as...
- Use
- Behavioral trajectory smoothing and binning module: As illustrated in C, the original behavioral trajectory for each frame often contains significant noise. For example, in a video where a mouse struggles for the first 6 s and remains immobile for the following 6 s, initial recognition may produce noisy results, as...
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Data and code availability
Any additional information required to re-analyze the data reported in this study is available from the upon request.
Behavioral trajectories in the FST and TST
To induce depression-like behaviors, we employed two experimental manipulations: chronic restraint stress ([CRS] A) and chemogenetic inhibition of the medial prefrontal cortex (mPFC) pyramidal neurons,,,, ( H). Average behavioral trajectories for CRS are presented in B and 3C, while those for mPFC inhibition are shown in I and 3J. Notably, the probabilities of swimming and struggling typically declined after test onset, with swimming showing a rebound primarily in the FST, whereas late-stage variability increased in both the FST and TST. In the FST, both CRS and mPFC inhibition led to reduced swimming behavior in the later stages compared to controls ( B and 3I). In the TST, CRS produced a reduction in struggling behavior during the early stages ( C). Conventional analyses of immobility times corroborate these findings and align with the existing literature ( D-3G and 3KR...
Behavioral trajectories in the FST and TST
(A) Experimental setup for CRS. Twenty-nine C57BL/6 mice were divided into two groups; the CRS group ( n = 13) underwent daily restraint stress (6-8 h/day) for 21 days, while controls ( n = 16) experienced no stress. Behavioral assessments were conducted on days 22 (FST) and 24 (TST).
Behavioral trajectories in the FST and TST
(D-G) Immobility times for the FST (D and E) and TST (F and G) in the CRS experiment are shown for the entire testing period (D and F) and the final 4 min (E and G). Blue and orange bars represent the control and CRS group, respectively, while black dots indicate individual animal data. Error bars denote the SEM.
Behaviors in the FST and TST are driven by different reinforcement learning processes
(C-F) Model-predicted vs. observed proportions of immobility (CRS experiment). Predictions for the (C and D) FST and (E and F) TST. Each pair compares model predictions with observed data over the full testing period (C and E) and the final 4 min (D and F). Colored bars indicate median predictions from the winning models, with error bars denoting the 95% highest density interval (HDI). Red dots present the observed group means, showing close alignment between model predictions and empirical data.
Behaviors in the FST and TST are driven by different reinforcement learning processes
To evaluate the predictive power of these winning models, we conducted posterior predictive checks. As shown in C-4F and 4M-4P, both models successfully captured the observed immobility proportions across all groups, both over the entire task and during the final 4 min. Nearly all individual immobility times fell within the predicted 95% highest density interval ( G-4J and 4Q-4T), with strong correlations between predicted and observed immobility times across all experimental conditions and time windows (r > 0.95, p < 10 -5, Pearson correlation). K, 4L, 4U, and 4V demonstrate that the models reliably predicted each mouse's latency to immobility (r > 0.71, p < 0.001, Pearson correlation). Moreover, the winning models accurately replicated the mice's behavioral trajectories ( W-4Z), capturing the dynamic evolution of behavior over time. C...
CRS and mPFC inhibition increase immobility by altering learning and consequence sensitivity
Within the framework of reinforcement learning, model parameters represent distinct cognitive components, thereby clarifying how specific factors contribute to increased immobility. In the FST ( A and 5B), both the CRS and hM4D groups demonstrated significant reductions in the learning rate (α) and the lower bound of consequence sensitivity adaptation (lb), alongside an elevated adaptation rate (β), compared to the controls. Additionally, the hM4D group exhibited a higher inverse temperature (τ). In the TST ( C and 5D), both CRS and hM4D groups displayed lower τ values than controls, while the CRS group further demonstrated increased scaling factor (κ) and attention updating rate (η), together with decreased consequence sensitivity (ρ). Figure 5 Uncovering cognitive processes driving immobility via model parameters
CRS and mPFC inhibition increase immobility by altering learning and consequence sensitivity
(A and B) Parameter distributions of the RW + ada model for the FST. This model comprises five parameters: the learning rate (α), which determines how rapidly animals update expectations; the inverse temperature (τ), which regulates the impact of expected values on decision-making; the lower bound of adaptation (lb), reflecting intensified negative experiences from repeated failed escape attempts; the upper bound of adaptation (ub), representing diminished negative perception after sustained immobility; and the adaptation rate (β), which dictates the speed of adaptation. Individual data points representing the mean of 2,000 samples per mouse, with group-level medians and interquartile ranges depicted by boxplots. Control, CRS, and hM4D groups are represented in blue, orange, and magenta, respectively.
Measurement outputs
What raw and processed outputs should exist?
(H) Chemogenetic inhibition of mPFC pyramidal neuron. Immunofluorescence confirmed hM4D(Gi)-mCherry expression in mPFC pyramidal neurons (red = hM4D(Gi), green = CaMKIIα, b...
- Raw artifact
- Field or section images captured from matched samples
- Processed artifact
- Selected representative panels with quantified intensity, counts, or area measurements
- Reported as
- Per-group imaging summaries with representative figures and quantified endpoints
To induce depression-like behaviors, we employed two experimental manipulations: chronic restraint stress ([CRS] A) and chemogenetic inhibition of the medial prefrontal cortex (...
- Raw artifact
- Per-sample or per-animal endpoint measurements collected during the experiment
- Processed artifact
- Structured table with cleaned measurements ready for comparison
- Reported as
- Summary statistics and between-group or across-timepoint comparisons
These detailed behavioral trajectories provide a richer dataset for nuanced analysis, as their dynamic patterns likely reflect underlying cognitive processes in mice. Building u...
- Raw artifact
- Per-sample or per-animal endpoint measurements collected during the experiment
- Processed artifact
- Structured table with cleaned measurements ready for comparison
- Reported as
- Summary statistics and between-group or across-timepoint comparisons
The observed behavioral trajectories exhibit patterns consistent with adaptive behavior, motivating the use of reinforcement learning as a computational framework to model these...
- Raw artifact
- Per-sample or per-animal endpoint measurements collected during the experiment
- Processed artifact
- Structured table with cleaned measurements ready for comparison
- Reported as
- Summary statistics and between-group or across-timepoint comparisons
Analysis plan
How should the outputs become interpretable results?
Acquisition
Collect raw experimental outputs with enough metadata to preserve sample identity, condition, and timing.
inferred from protocolPreprocessing / cleaning
(D-G) Immobility times for the FST (D and E) and TST (F and G) in the CRS experiment are shown for the entire testing period (D and F) and the final 4 min (E and G).
from paperScoring or quantification
Quantify the primary readouts for this experiment: (H) Chemogenetic inhibition of mPFC pyramidal neuron. Immunofluorescence confirmed hM4D(Gi)-mCherry expression in mPFC pyramidal neurons (red = hM4D(Gi), green = CaMKIIα, b...; To induce depression-like behaviors, we employed two experimental manipulations: chronic restraint stress ([CRS] A) and chemogenetic inhibition of the medial prefrontal cortex (...; These detailed behavioral trajectories provide a richer dataset for nuanced analysis, as their dynamic patterns likely reflect underlying cognitive processes in mice. Building u...; The observed behavioral trajectories exhibit patterns consistent with adaptive behavior, motivating the use of reinforcement learning as a computational framework to model these....
from paperStatistical comparison
(D-G) Immobility times for the FST (D and E) and TST (F and G) in the CRS experiment are shown for the entire testing period (D and F) and the final 4 min (E and G). Blue...; (K-N) Immobility times for the FST (G and H) and TST (I and L) in the mPFC inhibition experiment follow the same format as (D-G). Blue and magenta bars represent the...; Within the framework of reinforcement learning, model parameters represent distinct cognitive components, thereby clarifying how specific factors contribute to increased immobil...; (I-L) Predicted dynamics of expected value and attention allocation in the TST. (I and J) Struggling vs. immobility under CRS conditions; (K and L) predictions for mPFC in...
from paperReporting output
Report representative outputs alongside summary comparisons for (H) Chemogenetic inhibition of mPFC pyramidal neuron. Immunofluorescence confirmed hM4D(Gi)-mCherry expression in mPFC pyramidal neurons (red = hM4D(Gi), green = CaMKIIα, b..., To induce depression-like behaviors, we employed two experimental manipulations: chronic restraint stress ([CRS] A) and chemogenetic inhibition of the medial prefrontal cortex (..., These detailed behavioral trajectories provide a richer dataset for nuanced analysis, as their dynamic patterns likely reflect underlying cognitive processes in mice. Building u..., The observed behavioral trajectories exhibit patterns consistent with adaptive behavior, motivating the use of reinforcement learning as a computational framework to model these....
inferred from protocolStructured statistical methods
(D-G) Immobility times for the FST (D and E) and TST (F and G) in the CRS experiment are shown for the entire testing period (D and F) and the final 4 min (E and G). Blue...; (K-N) Immobility times for the FST (G and H) and TST (I and L) in the mPFC inhibition experiment follow the same format as (D-G). Blue and magenta bars represent the...; Within the framework of reinforcement learning, model parameters represent distinct cognitive components, thereby clarifying how specific factors contribute to increased immobil...; (I-L) Predicted dynamics of expected value and attention allocation in the TST. (I and J) Struggling vs. immobility under CRS conditions; (K and L) predictions for mPFC in...
source structuredSource and audit
What supports the facts on this page?
Evidence quotes (8)
Any additional information required to re-analyze the data reported in this study is available from the upon request.
To induce depression-like behaviors, we employed two experimental manipulations: chronic restraint stress ([CRS] A) and chemogenetic inhibition of the medial prefrontal cortex (mPFC) pyramidal neurons,,,, ( H). Average behavioral trajectories for CRS are presented in B and 3C, while those for mPFC inhibition are shown in I and 3J. Notably, the probabilities of swimming and struggling typically declined after test onset, with swimming showing a rebound primarily in the FST, whereas late-stage variability increased in both the FST and TST. In the FST, both CRS and mPFC inhibition led to reduced swimming behavior in the later stages compared to controls ( B and 3I). In the TST, CRS produced a reduction in struggling behavior during the early stages ( C). Conventional analyses of immobility times corroborate these findings and align with the existing literature ( D-3G and 3K-3N), supporting the reliability of our behavioral trajectory data. Furthermore, the CRS group exhibited significantly shorter inter-struggle intervals in both the FST ( E) and TST ( G). In the TST, CRS also reduced struggle bout duration ( K), with a similar trend observed in the FST ( I). By con...
(A) Experimental setup for CRS. Twenty-nine C57BL/6 mice were divided into two groups; the CRS group ( n = 13) underwent daily restraint stress (6-8 h/day) for 21 days, while controls ( n = 16) experienced no stress. Behavioral assessments were conducted on days 22 (FST) and 24 (TST).
(D-G) Immobility times for the FST (D and E) and TST (F and G) in the CRS experiment are shown for the entire testing period (D and F) and the final 4 min (E and G). Blue and orange bars represent the control and CRS group, respectively, while black dots indicate individual animal data. Error bars denote the SEM.
(C-F) Model-predicted vs. observed proportions of immobility (CRS experiment). Predictions for the (C and D) FST and (E and F) TST. Each pair compares model predictions with observed data over the full testing period (C and E) and the final 4 min (D and F). Colored bars indicate median predictions from the winning models, with error bars denoting the 95% highest density interval (HDI). Red dots present the observed group means, showing close alignment between model predictions and empirical data.
To evaluate the predictive power of these winning models, we conducted posterior predictive checks. As shown in C-4F and 4M-4P, both models successfully captured the observed immobility proportions across all groups, both over the entire task and during the final 4 min. Nearly all individual immobility times fell within the predicted 95% highest density interval ( G-4J and 4Q-4T), with strong correlations between predicted and observed immobility times across all experimental conditions and time windows (r > 0.95, p < 10 -5, Pearson correlation). K, 4L, 4U, and 4V demonstrate that the models reliably predicted each mouse's latency to immobility (r > 0.71, p < 0.001, Pearson correlation). Moreover, the winning models accurately replicated the mice's behavioral trajectories ( W-4Z), capturing the dynamic evolution of behavior over time. Collectively, these results confirm that the winning models robustly simulate mouse behavior.
Within the framework of reinforcement learning, model parameters represent distinct cognitive components, thereby clarifying how specific factors contribute to increased immobility. In the FST ( A and 5B), both the CRS and hM4D groups demonstrated significant reductions in the learning rate (α) and the lower bound of consequence sensitivity adaptation (lb), alongside an elevated adaptation rate (β), compared to the controls. Additionally, the hM4D group exhibited a higher inverse temperature (τ). In the TST ( C and 5D), both CRS and hM4D groups displayed lower τ values than controls, while the CRS group further demonstrated increased scaling factor (κ) and attention updating rate (η), together with decreased consequence sensitivity (ρ). Figure 5 Uncovering cognitive processes driving immobility via model parameters
(A and B) Parameter distributions of the RW + ada model for the FST. This model comprises five parameters: the learning rate (α), which determines how rapidly animals update expectations; the inverse temperature (τ), which regulates the impact of expected values on decision-making; the lower bound of adaptation (lb), reflecting intensified negative experiences from repeated failed escape attempts; the upper bound of adaptation (ub), representing diminished negative perception after sustained immobility; and the adaptation rate (β), which dictates the speed of adaptation. Individual data points representing the mean of 2,000 samples per mouse, with group-level medians and interquartile ranges depicted by boxplots. Control, CRS, and hM4D groups are represented in blue, orange, and magenta, respectively.
Machine-readable layer
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