Social Interaction and Novelty Recognition Tests (Three-Chamber Test)

The three-chamber test was used to evaluate social behavior. The testing apparatus consists of a rectangular box with three chambers separated by two doors. Each of the outer chambers contains a cage enclosure. The mice were habituated to the testing box for 5 min prior to testing. Testing occurred in two sessions. In the first session, a new intruder mouse (Stranger1) was placed in one cage, while the other cage was left empty. The tested mouse was placed in the middle chamber and left to explore the three chambers for 10 min. The time spent in the Stanger1 chamber versus the empty cage is used to evaluate social interaction of the test subject. In the second session, another intruder mouse (Stranger2) was placed in the second cage along with the previous intruder mouse (Stranger1) still held in the first cage, and the test subject was allowed to freely explore both chambers for 10 min. The time spent in the Stanger2 versus Stranger1 chamber was used to evaluate the social memory of the tested animal.