Behavioral effects of a chronic envy-like stress paradigm in mice using an adjacent cage model methods
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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...
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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...
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