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Automation of Continuous Spontaneous Alternation to Increase the Throughput for In Vivo Screening of Cognitive Enhancers. Optimization of the Ethovision System for the Y-maze Test in Mice

Continuous spontaneous alternation in a symmetrical Ymaze is widely considered as a measure of spatial working memory in rodents, and is used as an entrance screening test for cognitive enhancers. The task consists of a single 8min session during which arm visits and sequence of arm entries are continuously recorded. Using EthoVision XT, we describe the implementation and validation of an automated scoring procedure for continuous spontaneous alternation in mice in a Y-maze. We optimized the design of the arena setting to avoid false arm entries, and developed an automated analysis program in order to extract from the EthoVision export file the sequence of arm entries and the number of alternations. We show that, after optimization, 100% agreement was reached between the automated output and human observers in terms of arm entries, sequence of entry and alternation percentage. We also demonstrate that several Y-mazes can be run in parallel without influencing mouse alternation behavior, and that the automated system is suitable for pharmacological testing.