Behavioral effects of a chronic envy-like stress paradigm in mice using an adjacent cage model methods
Aim. Evidence-backed execution summary for Behavioral effects of a chronic envy-like stress paradigm in mice using an adjacent cage model methods from Behavioral effects of a chronic envy-like stress paradigm in mice using an adjacent cage model.
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mouse
Subject model for the experiment.
- Use
- confirm full cohort details in the source paper
Behavioral tests
reagent used in the protocol.
- Use
- All behavioral tests were conducted in a dedicated testing room during the light phase of the light/dark cycle, between 10:00 and 17:00. The behavioral test battery was designed to assess not only affective and cognitive domains but also general physiological and stress-related parameters. In addition to anxiety-, d...
Behavioral tests
All behavioral tests were conducted in a dedicated testing room during the light phase of the light/dark cycle, between 10:00 and 17:00. The behavioral test battery was designed to assess not only affective and cognitive domains but also general physiological and stress-related parameters. In addition to anxiety-, d...
- Use
- All behavioral tests were conducted in a dedicated testing room during the light phase of the light/dark cycle, between 10:00 and 17:00. The behavioral test battery was designed to assess not only affective and cognitive domains but also general physiological and stress-related parameters. In addition to anxiety-, d...
Light/dark transition test
Anxiety-like behavior was evaluated using the light/dark transition test. The apparatus consisted of an acrylic box (22 × 44 × 40 cm) divided into two compartments connected by a small opening (5 × 5 cm). One chamber had white acrylic walls and was brightly illuminated (2...
- Use
- Anxiety-like behavior was evaluated using the light/dark transition test. The apparatus consisted of an acrylic box (22 × 44 × 40 cm) divided into two compartments connected by a small opening (5 × 5 cm). One chamber had white acrylic walls and was brightly illuminated (2...
Y-maze test
Spatial working memory was evaluated using the Y-maze test. The apparatus consisted of three identical white acrylic arms (length: 40 cm; lower arm width: 3 cm; upper arm width: 10 cm; wall height: 12 cm) arranged at 120° angles from each other. Each mouse was placed in the center of the...
- Use
- Spatial working memory was evaluated using the Y-maze test. The apparatus consisted of three identical white acrylic arms (length: 40 cm; lower arm width: 3 cm; upper arm width: 10 cm; wall height: 12 cm) arranged at 120° angles from each other. Each mouse was placed in the center of the...
Social interaction test in a novel environment
Social behavior was assessed using a social interaction test conducted in a novel environment. The apparatus consisted of a square white acrylic box (45 × 45 × 40 cm) illuminated at 100 lux. Two unfamiliar mice of the same sex, originating from different home cages, were simultaneously pl...
- Use
- Social behavior was assessed using a social interaction test conducted in a novel environment. The apparatus consisted of a square white acrylic box (45 × 45 × 40 cm) illuminated at 100 lux. Two unfamiliar mice of the same sex, originating from different home cages, were simultaneously pl...
Tail-suspension test
Depressive-like behavior was evaluated using the tail suspension test. Mice were suspended by the tail in a white plastic chamber positioned 60 cm above the floor, secured with adhesive tape placed less than 1 cm from the tip of the tail. Behavior was recorded for 6 min using a video camera, and im...
- Use
- Depressive-like behavior was evaluated using the tail suspension test. Mice were suspended by the tail in a white plastic chamber positioned 60 cm above the floor, secured with adhesive tape placed less than 1 cm from the tip of the tail. Behavior was recorded for 6 min using a video camera, and im...
Porsolt forced-swim test
Depressive-like behavior was further evaluated using the Porsolt forced swim test. The apparatus consisted of a Plexiglas cylinder (20 cm in height × 10 cm in diameter) filled with water maintained at 23 °C to a depth of 7.5 cm. Each mouse was placed in the cylinder for 6 min, and b...
- Use
- Depressive-like behavior was further evaluated using the Porsolt forced swim test. The apparatus consisted of a Plexiglas cylinder (20 cm in height × 10 cm in diameter) filled with water maintained at 23 °C to a depth of 7.5 cm. Each mouse was placed in the cylinder for 6 min, and b...
Statistical analysis
All data are expressed as box plots with individual data points. Statistical analyses were performed using GraphPad Prism (GraphPad Software, San Diego, CA, USA). For each behavioral parameter, comparisons between control and envy groups were made using two-tailed Student's t -tests. A p value of < 0.05...
- Use
- All data are expressed as box plots with individual data points. Statistical analyses were performed using GraphPad Prism (GraphPad Software, San Diego, CA, USA). For each behavioral parameter, comparisons between control and envy groups were made using two-tailed Student's t -tests. A p value of < 0.05...
Chronic adjacent-cage exposure designed to induce envy-like stress does not affect spatial working memory in the Y-ma...
We next evaluated spatial working memory using the Y-maze test ( ). The chronic exposure paradigm did not affect general locomotor activity, with no significant differences observed in the total distance traveled ( A-D ) or the total number of arm entries ( E-H ). Furthermore, spatial working memory rema...
- Use
- We next evaluated spatial working memory using the Y-maze test ( ). The chronic exposure paradigm did not affect general locomotor activity, with no significant differences observed in the total distance traveled ( A-D ) or the total number of arm entries ( E-H ). Furthermore, spatial working memory rema...
Statistical analysis
Software used for acquisition, scoring, statistics, or reporting.
- Use
- All data are expressed as box plots with individual data points. Statistical analyses were performed using GraphPad Prism (GraphPad Software, San Diego, CA, USA). For each behavioral parameter, comparisons between control and envy groups were made using two-tailed Student's t -tests. A p value of < 0.05...
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Methods
Male C57BL/6 N mice served as observers, while demonstrator mice were assigned to one of four conditions: (1) an environment enriched with objects, (2) an igloo, (3) a tube, or (4) social isolation. Observers were continuously exposed to these adjacent cages for 21 days. Subsequently, a comprehensive battery of behavioral tests was conducted to assess general health, anxiety-like behavior, spatial memory, social behavior, and depression-like behavior.
Animals
Eight-week-old male C57BL/6 N mice were purchased from The Jackson Laboratory Japan (Tokyo, Japan). A total of XX mice were used in this study. For each experimental condition (objects, isolation, igloos, and tubes), observer mice were randomly assigned to either the control group (n = 10) or the envy group (n = 10). Only male mice were used to avoid potential confounding effects of sex-dependent variability; however, this represents a limitation of the study. Mice were group-housed in transparent plastic cages (220 × 340 × 150 mm) with a nonwoven fabric filter cap. Standard laboratory chow (MF-R, Oriental Yeast Co., Tokyo, Japan), nesting material, and water were provided ad libitum. Animals were maintained in a temperature-controlled room (23-26 °C) under a 12-h light/dark cycle (lights on at 08:00, lights off at 2...
Envy-like stress paradigm
For the experimental design, each observation mouse was housed in a cage adjacent to the demonstrator cage under one of four environmental conditions (object, isolation, igloo, or tube). To minimize the effects of location, all cages in the group were placed on racks at the same height in the animal room. To avoid environmental influences such as odor, sound, and location, all behavioral experiments were conducted in a room separate from the animal care facility. The envy-like stress paradigm was designed to evaluate the effects of prolonged vicarious exposure to conspecifics housed under different environmental conditions. Each observer mouse was group-housed in a transparent plastic cage (220 × 340 × 150 mm) placed adjacent to a demonstrator cage of the same size. Observer mice were group-housed throughout the experiment to avoid confounding effects assoc...
Social interaction test in a novel environment
Social behavior was assessed using a social interaction test conducted in a novel environment. The apparatus consisted of a square white acrylic box (45 × 45 × 40 cm) illuminated at 100 lux. Two unfamiliar mice of the same sex, originating from different home cages, were simultaneously placed in opposite corners of the arena. Each pair was allowed to interact freely for 10 min. The parameters measured were total distance traveled (m), number of contacts, total contact time (s), and mean duration per contact (s). Behavioral data were analyzed using SMART software, version 3.0 (PanLab, Harvard Apparatus, Spain).
Observing a socially isolated demonstrator reduced the duration of social interactions in observer mice
To determine the effects of the exposure paradigm on social behavior, we used the social interaction test in a novel environment ( ). There were no significant differences between the control and envy groups in total distance traveled during the test ( A-D ). The overall amount of social interaction was also unaffected, with no significant changes in the number of social contacts ( E-H ) or the total time spent in social contact ( I-L ) in any of the four conditions. However, when examining the quality of these interactions, we found that observer mice in the isolation condition had a significantly shorter mean duration per contact compared to their controls ( N ). This effect was specific to the isolation condition, as no significant differences were found in the objects, igloos, or tubes conditions ( M, O, P ). Fig. 5 Social behavior of observer mice in a novel env...
Measurement outputs
What raw and processed outputs should exist?
Male C57BL/6 N mice served as observers, while demonstrator mice were assigned to one of four conditions: (1) an environment enriched with objects, (2) an igloo, (3) a tub...
- Raw artifact
- Per-run gait capture with paw placement, timing, and stride features for each animal
- Processed artifact
- Cleaned gait metrics table and recovery trend summary across timepoints
- Reported as
- Group comparisons of gait indices, stride metrics, or recovery curves
For the experimental design, each observation mouse was housed in a cage adjacent to the demonstrator cage under one of four environmental conditions (object, isolation, igloo,...
- Raw artifact
- Per-run gait capture with paw placement, timing, and stride features for each animal
- Processed artifact
- Cleaned gait metrics table and recovery trend summary across timepoints
- Reported as
- Group comparisons of gait indices, stride metrics, or recovery curves
All behavioral tests were conducted in a dedicated testing room during the light phase of the light/dark cycle, between 10:00 and 17:00. The behavioral test battery was designed...
- Raw artifact
- Per-run gait capture with paw placement, timing, and stride features for each animal
- Processed artifact
- Cleaned gait metrics table and recovery trend summary across timepoints
- Reported as
- Group comparisons of gait indices, stride metrics, or recovery curves
Neuromuscular strength was evaluated using a grip-strength meter (O'Hara & Co., Tokyo, Japan). Each mouse was gently lifted by the tail and allowed to grasp the wire mesh...
- Raw artifact
- Per-run gait capture with paw placement, timing, and stride features for each animal
- Processed artifact
- Cleaned gait metrics table and recovery trend summary across timepoints
- Reported as
- Group comparisons of gait indices, stride metrics, or recovery curves
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
All data are expressed as box plots with individual data points.
from paperScoring or quantification
Quantify the primary readouts for this experiment: Male C57BL/6 N mice served as observers, while demonstrator mice were assigned to one of four conditions: (1) an environment enriched with objects, (2) an igloo, (3) a tub...; For the experimental design, each observation mouse was housed in a cage adjacent to the demonstrator cage under one of four environmental conditions (object, isolation, igloo,...; All behavioral tests were conducted in a dedicated testing room during the light phase of the light/dark cycle, between 10:00 and 17:00. The behavioral test battery was designed...; Neuromuscular strength was evaluated using a grip-strength meter (O'Hara & Co., Tokyo, Japan). Each mouse was gently lifted by the tail and allowed to grasp the wire mesh....
from paperStatistical comparison
All data are expressed as box plots with individual data points. Statistical analyses were performed using GraphPad Prism (GraphPad Software, San Diego, CA, USA). For each behav...; To assess anxiety-like behavior, we subjected the mice to the light/dark transition test ( ). General locomotor activity, measured by the total distance traveled and the number...; We next evaluated spatial working memory using the Y-maze test ( ). The chronic exposure paradigm did not affect general locomotor activity, with no significant differences obse...; To determine the effects of the exposure paradigm on social behavior, we used the social interaction test in a novel environment ( ). There were no significant differences betwe...
from paperReporting output
Report representative outputs alongside summary comparisons for Male C57BL/6 N mice served as observers, while demonstrator mice were assigned to one of four conditions: (1) an environment enriched with objects, (2) an igloo, (3) a tub..., For the experimental design, each observation mouse was housed in a cage adjacent to the demonstrator cage under one of four environmental conditions (object, isolation, igloo,..., All behavioral tests were conducted in a dedicated testing room during the light phase of the light/dark cycle, between 10:00 and 17:00. The behavioral test battery was designed..., Neuromuscular strength was evaluated using a grip-strength meter (O'Hara & Co., Tokyo, Japan). Each mouse was gently lifted by the tail and allowed to grasp the wire mesh....
inferred from protocolStructured statistical methods
All data are expressed as box plots with individual data points. Statistical analyses were performed using GraphPad Prism (GraphPad Software, San Diego, CA, USA). For each behav...; To assess anxiety-like behavior, we subjected the mice to the light/dark transition test ( ). General locomotor activity, measured by the total distance traveled and the number...; We next evaluated spatial working memory using the Y-maze test ( ). The chronic exposure paradigm did not affect general locomotor activity, with no significant differences obse...; To determine the effects of the exposure paradigm on social behavior, we used the social interaction test in a novel environment ( ). There were no significant differences betwe...
source structuredSource and audit
What supports the facts on this page?
Evidence quotes (5)
Male C57BL/6 N mice served as observers, while demonstrator mice were assigned to one of four conditions: (1) an environment enriched with objects, (2) an igloo, (3) a tube, or (4) social isolation. Observers were continuously exposed to these adjacent cages for 21 days. Subsequently, a comprehensive battery of behavioral tests was conducted to assess general health, anxiety-like behavior, spatial memory, social behavior, and depression-like behavior.
Eight-week-old male C57BL/6 N mice were purchased from The Jackson Laboratory Japan (Tokyo, Japan). A total of XX mice were used in this study. For each experimental condition (objects, isolation, igloos, and tubes), observer mice were randomly assigned to either the control group (n = 10) or the envy group (n = 10). Only male mice were used to avoid potential confounding effects of sex-dependent variability; however, this represents a limitation of the study. Mice were group-housed in transparent plastic cages (220 × 340 × 150 mm) with a nonwoven fabric filter cap. Standard laboratory chow (MF-R, Oriental Yeast Co., Tokyo, Japan), nesting material, and water were provided ad libitum. Animals were maintained in a temperature-controlled room (23-26 °C) under a 12-h light/dark cycle (lights on at 08:00, lights off at 20:00). All mice were acclimated to these conditions for at least one week before the start of any experimental procedures. Cages were cleaned once per week.
For the experimental design, each observation mouse was housed in a cage adjacent to the demonstrator cage under one of four environmental conditions (object, isolation, igloo, or tube). To minimize the effects of location, all cages in the group were placed on racks at the same height in the animal room. To avoid environmental influences such as odor, sound, and location, all behavioral experiments were conducted in a room separate from the animal care facility. The envy-like stress paradigm was designed to evaluate the effects of prolonged vicarious exposure to conspecifics housed under different environmental conditions. Each observer mouse was group-housed in a transparent plastic cage (220 × 340 × 150 mm) placed adjacent to a demonstrator cage of the same size. Observer mice were group-housed throughout the experiment to avoid confounding effects associated with social isolation. However, group housing may have provided social buffering, which could have mitigated stress-related and envy-like behavioral effects. A transparent wall allowed visual access between cages, while airflow through the filter caps ensured the presence of olfactory cues. De...
Social behavior was assessed using a social interaction test conducted in a novel environment. The apparatus consisted of a square white acrylic box (45 × 45 × 40 cm) illuminated at 100 lux. Two unfamiliar mice of the same sex, originating from different home cages, were simultaneously placed in opposite corners of the arena. Each pair was allowed to interact freely for 10 min. The parameters measured were total distance traveled (m), number of contacts, total contact time (s), and mean duration per contact (s). Behavioral data were analyzed using SMART software, version 3.0 (PanLab, Harvard Apparatus, Spain).
To determine the effects of the exposure paradigm on social behavior, we used the social interaction test in a novel environment ( ). There were no significant differences between the control and envy groups in total distance traveled during the test ( A-D ). The overall amount of social interaction was also unaffected, with no significant changes in the number of social contacts ( E-H ) or the total time spent in social contact ( I-L ) in any of the four conditions. However, when examining the quality of these interactions, we found that observer mice in the isolation condition had a significantly shorter mean duration per contact compared to their controls ( N ). This effect was specific to the isolation condition, as no significant differences were found in the objects, igloos, or tubes conditions ( M, O, P ). Fig. 5 Social behavior of observer mice in a novel environment after chronic adjacent-cage exposure designed to induce envy-like stress. Social behavior in a novel environment was assessed by ( A-D ) total distance traveled, ( E-H ) number of social contacts, ( I-L ) total duration of social interaction, and ( M-P ) mean duratio...
Machine-readable layer
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