Individual housing and handling procedures modify anxiety levels of Tg2576 mice assessed in the zero maze test

The zero maze is an unconditioned anxiety test for mice, in which a number of environmental variables can modify the anxiety levels of the animals. In the present study, we have assessed how individual housing, handling procedure and interaction between individual housing and handling procedure affect the baseline anxiety of mice. Thirty-seven wild type mice and eighteen Tg2576 mice were used (obtained from crossing APPSWE hemizygous male C57BL6/SJL background with C57BL6/SJL female). Wild type mice were randomly assigned to four experimental groups: 1) group housed and unhandled, 2) group housed but handled, 3) individually housed, unhandled, and 4) individually housed and handled. In turn, Tg2576 mice were randomly assigned to two experimental groups: 1) individually housed, unhandled, and 2) individually housed and handled. The results show that individually housed mice exhibited more anxiety-related behaviors over a 5 min testing period than the other experimental groups. Use of the handling procedure was associated with a statistically significant reduction in anxiety-related behaviors among individually housed mice. No effects on anxiety-related behavior levels were observed when group housed animals were handled. When activity levels were significantly increased, a new parameter, “ Time by Entries ”, helped to prevent activity from influencing anxiety parameters such as time in the open section of the zero maze test. This knowledge can help to design more efficient experiments without bias from data obtained by means of unconditioned tests.