Elevated Plus Maze
Objective: Assessment of anxiety-like behavior in mice using the elevated plus maze apparatus
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Protocol Steps
Animal housing and maintenance
Littermate mice housed in temperature-controlled and light-controlled pathogen-free room with ad libitum access to water and standard regular chow
Note: Light cycles: 12 h with lights on at 0700 h. Housing in Green Line IVC Sealsafe PLUS cages (Tecniplast)
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”
Behavioral testing room habituation
Animals given habituation period to the behavioral testing room prior to testing
Note: Habituation performed before behavioral phenotyping
View evidence from paper
“Animals were given 30 min of habituation to the behavioral testing room”
Elevated plus maze testing
Elevated plus maze test performed as part of behavioral phenotyping battery. Test order determined by invasiveness level to minimize prior test history influence
Note: Performed between 0900 h and 1600 h. Elevated plus maze is the second test in the sequence (after rotarod, before marble burying test). 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. Tests were performed from the least to the most invasive to minimize the influence of prior test history (in order: rotarod, elevated plus maze, marble burying test, open field test, sucrose preference test, novelty suppressed feeding and forced swim test)”
Chronic restraint stress (if applicable)
After baseline behavioral testing, animals submitted to restraint stress for 21 days. Mice placed in horizontal resting position inside well-ventilated 50 ml falcon tube daily
Note: Restraint stress applied at 1000 h each day. Tube ventilated with 12 holes of 0.5 mm diameter. Test order reversed after chronic restraint stress.
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”