Spontaneous Alternation Test (Y-Maze)

Spontaneous alternation is another test of spatial memory and was performed using a Y-maze with symmetrical arms. Each arm of the maze was 60 cm long, 10 cm wide, and 15 cm high.⁵¹ This test consisted of only one 5-minute trial and was conducted in a sound-insulated room. The mice were individually placed at the end of one arm and allowed to explore all arms of the maze freely. Between each trial, the maze arms were wiped with 70% alcohol to avoid odour cues.⁵² The maze arms were labelled A, B, and C. The spontaneous alternation score was defined as sequential entry into the three different arms (ie, ABC, CBA, or BAC). Re-entry into the same arm such as ABA or ACA was considered an entry error. The spontaneous alternation percentage (SAP) was calculated by SAP = number of alternations/(total arm entries – 2) × 100.⁵¹