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Phenotyping

Y-maze spontaneous alternation

Purpose

This test allows evaluation of working memory. When placed in the Y-maze, mice tend to alternate visits between the three arms. A mouse with impaired working memory cannot remember which arm it just visited and thus shows decreased spontaneous alternation.

Equipment

Home-made equipment made of PVC.

Recommendations

10 mice per group are recommended for reliable data analysis.

Pavlovian fear conditioning

Purpose

This is an associative learning paradigm for measuring emotional learning and memory. A neutral conditioned stimulus (CS, tone) is paired with an aversive unconditioned stimulus (US, foot-shock). After conditioning, the CS or the spatial context alone elicits a central state of fear expressed as reduced locomotor activity or total lack of movement (freezing).

Equipment

4 automated cages (Coulbourn Instruments, Whitehall, USA)

Recommendations

10 mice per group are recommended for reliable data analysis.

Object recognition

Purpose

The object recognition task is based on the natural tendency of rodents to explore a novel object / environment in comparison to a familiar one. This test allows evaluation of recognition memory to characterize potential memory defects. 

Equipment

Automated openfield arenas (Panlab, Barcelone, Spain) and different objects (marble, dice…) are required. 

Recommendations

10 mice per group are recommended for reliable data analysis.

Morris water maze

Purpose

This test is used to evaluate spatial reference memory in rodents. Mice are trained to escape from water by swimming to a submerged platform using only distal extra-maze cues.

Equipment:

2 rooms dedicated to water maze test and videotracking systems are used.

Recommendations

10 mice per group are recommended for reliable data analysis.

Active/passive avoidance

Purpose

These tests are used for evaluation of emotional learning and memory processes. In the active avoidance test, a conditioned stimulus (CS, light/tone) is paired with an unconditioned stimulus (US, aversive foot-shock).  The animals can avoid the foot-shock actively by leaving the compartment where they are in the interval-time between the CS onset and the US occurrence. In the passive task, mice receive a foot shock in a dark compartment. After conditioning, they can passively avoid, by staying in the light side where they are placed.

Equipment

4 automated cages (Panlab, barcelona, Spain).

Recommendations

10 mice per group are recommended for reliable data analysis.

Scopolamine-induced amnesia

Purpose

The scopolamine model is based on the fact that cholinergic system is altered in amnesic syndromes like Alzheimer's disease. Administration of scopolamine cause memory deficits in several tests.

Equipment

Any equipment needed in tests used for evaluation of learning and memory.

Recommendations

10 mice per group are recommended for reliable data analysis.

TouchScreen

Purpose

Touchscreen technology allows to explore cognitive deficits with a platform that is translatable across rodents, non-human primates and human subjects (Palmer et al., 2021) lending itself to a high degree of standardization and throughput.This methodology is low stress, using appetitive rather than aversive reinforcement.

Sample test results

In addition to the traditional behavioral tests used to study cognitive skills, such as novel object recognition, Morris water maze, and fear conditioning, our team has implemented a touchscreen pipeline to assess visual discrimination, cognitive flexibility, and spatial learning.

After pre-training to learn to press an image that appears on the screen in order to obtain a reward of sweet water, the mice are tested in 3 different protocols.

  • In the pairwise discrimination, one of two shapes displayed simultaneously on the screen is correct. Touching the correct shape will be rewarded with sweet liquid. Touching the incorrect shape will be punished with a timeout. This simple task allows to evaluate the visual discrimination of animals.
  • Once the task has been learnt, the stimuli are reversed so that the correct shape now becomes the incorrect shape and vice versa. This reversal learning requires inhibition of prepotent responses and is known to be dependent on the prefrontal cortex.
  • Finally, mice are tested in the paired associates learning, a hippocampus-dependent task. Two flowers or two airplanes are presented, with the left flower and the right airplane used as the correct stimuli rewarded with sweet liquid.

Equipment

12 Mouse Touch Screen Systems (Campden Instruments)
Strawberry syrup

Recommendations

12 mice per group are recommended for reliable data analysis.
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