Chronic Restraint Stress
Objective: To expose mice to chronic physical restraint stress for 21 days to induce stress-related behavioral and physiological changes
This is a Chronic Restraint Stress protocol using mouse as the model organism. The procedure involves 3 procedural steps, 3 materials. Extracted from a 2016 paper published in Molecular Psychiatry.
Model and subjects
mouse • C57BL/6J background • male • 60-90 days • 101
Study window
~12.9 week study window | ~30 minutes hands-on
Core workflow
Animal housing and acclimation • Baseline behavioral testing • Chronic restraint stress exposure
Primary readouts
- Behavioral responses measured by rotarod, elevated plus maze, marble burying test, open field test, sucrose preference test, novelty suppressed feeding, and forced swim test
Key equipment and reagents
Use this page as an execution guide, then fall back to the source paper whenever you need exact exclusions, dosing details, or assay-specific caveats.
Confirm first
- Verify the animal model, intervention setup, and collection timepoints against the source paper.
- Check that every direct vendor link matches the exact specification your lab plans to run.
Use the page like this
- Work through the protocol steps in order and use the inline vendor chips only when you need to source or verify an item.
- Jump to Experimental Context for readouts, data shape, and analysis flow before planning downstream analysis.
Protocol Steps
Start here. The step list is optimized for running the experiment, with direct vendor links available inline when you need to source a cited item.
Animal housing and acclimation
Male mice aged 60-90 days were group housed in individually ventilated cages in a pathogen-free room with controlled temperature and light cycles
Note: Light cycle: 12 h cycles, lights on at 0700 h; water and standard regular chow provided ad libitum
View evidence from paper
“Littermates were group housed (Green Line IVC Sealsafe PLUS mouse, Tecniplast, Varese, Italy) in a temperature-specific (22C±1°C) and light-specific (12 h cycles, lights on at 0700 h) pathogen-free room with water and standard regular chow ad libitum”
Baseline behavioral testing
Animals underwent behavioral phenotyping prior to chronic restraint stress exposure. Tests performed in order from least to most invasive: rotarod, elevated plus maze, marble burying test, open field test, sucrose preference test, novelty suppressed feeding, and forced swim test
Note: Investigators were not blinded to group assignment; test order reversed after chronic restraint stress for bell-shaped stress exposure
View evidence from paper
“Behavioral phenotyping was performed between 0900 h and 1600 h. Animals were given 30 min of habituation to the behavioral testing room. Tests were performed from the least to the most invasive”
Chronic restraint stress exposure
After baseline behavioral testing, animals were placed in a horizontal resting position inside a well-ventilated 50 ml falcon tube for restraint stress
Note: Restraint tubes are well-ventilated with 12 holes of 0.5 mm diameter; restraint performed daily at 1000 h
View evidence from paper
“After baseline behavioral testing, animals were submitted to restraint stress for 21 days. Every day, mice were placed in a horizontal resting position inside a well-ventilated (12 holes, 0.5 mm diameter) 50 ml falcon tube at 1000 h and after 4-6 h they were unrestrained”