2.2.5 Three-Chamber Test (TCT)
Social behaviour of the mice was evaluated by the TCT following a
previously described protocol (Companys-Alemany et al., 2020). A box
(15x15x20 com) divided into three equally dimensioned rooms with
openings among them was used. The mice were submitted to 15-min trials.
First, each mouse was placed in the centre of the box and allowed to
explore the three chambers for 5 min (habituation). The entries to each
room were measured manually. Afterwards, an intruder (same-sex and age)
was placed in a metal cage at one of the rooms, and behaviour was
recorded for 10 min. The time spent in each room and the time
interacting with the intruder (e.g., sniffing, grooming) were measured
manually. The TCT apparatus was cleaned with 70% ethanol between the
trials to eliminate olfactory cues.