Several behavioral assays are currently used for high-throughput neurophenotyping and screening of genetic mutations and psychotropic drugs in zebrafish (Danio rerio). In this protocol, we describe a battery of two assays to characterize anxiety-related behavioral and endocrine phenotypes in adult zebrafish. Here, we detail how to use the 'novel tank' test to assess behavioral indices of anxiety (including reduced exploration, increased freezing behavior and erratic movement), which are quantifiable using manual registration and computer-aided video-tracking analyses. In addition, we describe how to analyze whole-body zebrafish cortisol concentrations that correspond to their behavior in the novel tank test. This protocol is an easy, inexpensive and effective alternative to other methods of measuring stress responses in zebrafish, thus enabling the rapid acquisition and analysis of large amounts of data. As will be shown here, fish anxiety-like behavior can be either attenuated or exaggerated depending on stress or drug exposure, with cortisol levels generally expected to parallel anxiety behaviors. This protocol can be completed over the course of 2 d, with a variable testing duration depending on the number of fish used.

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The zebrafish (Danio rerio) is becoming increasingly popular in the field of neurobehavioral research, including experimental, genetic, and pharmacological models of human brain disorders. While zebrafish research is rapidly expanding, its application as a translational neurobehavioral model is still in its relative infancy. Therefore, further investigation of new models is needed for targeting more domains and new, more complex brain disorders. The main aim of this paper is to discuss recent developments in the field of zebrafish neurobehavioral research, and to outline important emerging topics for further studies.