Elevated cross maze

Summary

The experiment was carried out on a "+" shaped device with two open and two closed arms, each of which was 10 x 50 cm long and 55 cm high. placing the animals in it meant that the animals were confronted with a struggle between the tendency to explore new environments and the fear of heights and open spaces, and thus a similar state of anxiety was characterized by increased avoidance of the open arms compared to the normal animals. state of anxiety was characterized by increased avoidance of the open arms.

Operation method

Elevated Plus Maze Test (Elevated Plus Maze Test)

Principle

The laboratory was conducted on a "+" shaped device with two open arms and two closed arms, each 10 x 50 cm long and 55 cm high. placing the animals in it meant that the animals were faced with a struggle between the tendency to explore new environments and the fear of heights and open spaces, so that compared to normal animals, a state similar to anxiety was characterized by increased avoidance of open arms. Anxiety-like state is characterized by increased avoidance in the open arms.

Materials and Instruments

Alcohol spritzers, kitchen paper, elevated cross maze, camera tracking system.

Move

1. The maze should be assembled in an isolated room away from any external disturbances from noise, odors, or human movement. Optionally, a low-intensity white noise source can be placed in the behavior lab. It should be noted that guiding cues, such as drawings on the walls around the room, may skew the animal's activity to a particular area of the maze. Therefore, any paintings or artwork that may serve as a cue must be removed.

2. Throughout the experiment, the experimenter must restrain himself from making too much noise or moving around, and refrain from using perfume, cologne, or any product with a strong odor, as it may produce an anxiety stimulus in the mice.

3. The illuminance in the room must be measured with the help of an illuminance meter, kept constant and controlled according to the analysis to be performed. Considering that low-intensity brightness reduces open-arm avoidance, in order to analyze anxiety effects, low-intensity illumination (5-30 Lux) should be preferred, while anxiolytic effects should be analyzed under high-intensity illumination (200-400 Lux or higher).

4. After the above conditions are met, the animals are placed in the experimental room for at least 45-60 minutes to acclimatize to the environment.

5. Before starting the test, clean the labyrinth with 70% ethanol to remove any dirt or odors that may have accumulated on the apparatus.

6. Turn on the video camera and place the first mouse in the center of the maze with the mouse facing an open arm (preferably the one opposite the experimenter).

7. Allow the animal to explore freely for 5 minutes and then remove the animal.

8. All urine and feces must be removed and the maze thoroughly cleaned with 70% ethanol to remove any residual odor from the first mouse. Then, switch to the next animal.

Image cited in:
Leo, L. M. and Pamplona, F. A. (2014). Elevated Plus Maze Test to Assess Anxiety-like Behavior in the Mouse. bio-protocol 4(16): e1211. doi: 10.21769/BioProtoc.1211.

Caveat

1. The tester tries to stay away from the animal's line of sight during the test to prevent artificially caused interference with the results.

2. When each animal is alternating, the maze must be wiped clean of odor and feces with alcohol.

Common Problems

The results of the experiment always jump?

A: Behavioral experiments have high requirements for the environment, especially the elevated cross maze this kind of experiments to detect anxiety, slight environmental changes can cause anxiety behavior of animals, so before doing the experiment, it is best that the tester can handle the animal in advance, pet the animal, so that the animal will not be anxious because of the tester, the next step is the preparation of the environment, which must be clean and odorless.

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