4.3.3. Three-Chamber Social Interaction Test (Crawley’s Sociability and Preference for Social Novelty Test)

The three-chamber social interaction test was performed on forty mice (20 in each experimental group), as described previously with modifications [33]. This test assesses general sociability in rodents. The mouse normally prefers to spend more time with another mouse. At PND 58–59, mice were introduced to a three-chamber social interaction apparatus (100 × 55 × 30 cm). Openings between the compartments (10 cm-wide doors) allowed the animals to access all three chambers. In phase I, each tested animal was allowed to explore the environment freely for 10 min for habituation. After the habituation phase (phase I), the subject was gently guided by the operator to the central chamber, and the two entrances were blocked. Two metal wire cages, the first one containing a sex-, age-, and weight-matched mouse and the second one empty, were placed in the left and right chambers (the order was randomised). Then, the two entrances were opened to allow the tested animal to explore the new environment freely for 10 min, and the social preference was measured (social stimulus vs. non-social stimulus)—phase II. Individual movement tracks were recorded by using a video system and analysed using the BehaView 0.0.16 software http://www.pmbogusz.net/?a=behaview (accessed on 24 August 2021). The time spent in each chamber and the time spent on direct interaction with the animal were measured by a blinded operator.