Background A dearth of laboratory tests to study actual human approach-avoidance behavior has complicated translational research on anxiety. The elevated plus-maze (EPM) is the gold standard to assess approach-avoidance behavior in rodents. Methods Here, we translated the EPM to humans using mixed reality through a combination of virtual and real-world elements. In two validation studies, we observed participants’ anxiety on a behavioral, physiological, and subjective level. Results Participants reported higher anxiety on open arms, avoided open arms, and showed an activation of endogenous stress systems. Participants’ with high anxiety exhibited higher avoidance. Moreover, open arm avoidance was moderately predicted by participants’ acrophobia and sensation seeking, with opposing influences. In a randomized, double blind, placebo controlled experiment, GABAergic stimulation decreased avoidance of open arms while alpha-2-adrenergic antagonism increased avoidance. Conclusion These findings demonstrate cross-species validity of open arm avoidance as a translational measure of anxiety. We thus introduce the first ecologically valid assay to track actual human approach-avoidance behavior under laboratory conditions.

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