Three-dimensional reconstruction is a method of qualifying the behavioral activity of several animals including mice, rats, and zebrafish. This method allows for measuring behavioral endpoint data on two types of tracking planes (temporal and spatial). Temporal tracking measures the activity fa subject across time while spatial tracking measures the activity of a subject in a specific area of the experimental arena as such. Data representation over 3D visual trajectory reconstruction is a robustly advantageous method of behavioral phenotyping. Digital video-tracking and analysis eliminates the inaccuracies of manual tracking and allows for improved investigation of behavioral activity at specific points in time or specific areas of the tracking arena.

3,4-Methylenedioxymethamphetamine (MDMA, ‘ecstasy’) is a potent psychedelic drug inducing euphoria and hypersociability in humans, as well as hyperactivity and anxiety in rodents. Adult zebrafish (Danio rerio) have become a widely used species in neurobehavioral research. Here, we explore the effects of a wide range (0.25–120 mg/l) of acute MDMA doses on zebrafish behavior in the novel tank test. Although MDMA was inactive at lower doses (0.25–10 mg/l), higher doses reduced bottom swimming and immobility (40–120 mg/l) and impaired intrasession habituation (10–120 mg/l). MDMA also elevated brain c-fos expression, collectively confirming the usage of zebrafish models for screening of hallucinogenic compounds.

Here, we integrated manual observations, computerized event‐based scoring and automated video analysis of zebrafish traces to generate three‐dimensional (3D) swim path reconstruction of general behavioral patterns, thereby improving the characterization and interpretation of complex, anxiety‐related adult zebrafish behavior. Drugs known to evoke a high‐anxiety state (caffeine, repeated morphine withdrawal) or an anxiolytic‐like effect (ethanol, morphine, fluoxetine, nicotine) wereused to detect multiple anxiety‐like behaviors and correlate them to those established manual observations. Additionally, we used this method to characterize the effects of lysergic acid diethylamide and 3,4‐methylenedioxymethamphetamine on zebrafish novelty‐evoked exploration. Our approach markedly enhances the power of zebrafish behavioral pharmacology, by providing innovative techniques for high‐throughput, multidimensional neurophenotyping of adult zebrafish behavior.

Ketamine is a non-competitive glutamatergic antagonist used to induce sedation and analgesia. In sub-anesthetic doses, it induces hyperlocomotion, impairs memory and evokes stereotypic circling in rodents. Zebrafish (Danio rerio) emerged as a promising new animal model to screen the effects of psychotropic compounds. Here, we investigated the effects of sub-anesthetic doses of ketamine on anxiety, locomotion, habituation and social behavior of adult zebrafish. Acute 20-min exposure to 20 and 40 mg/L (but not 2 mg/L) of ketamine reduced anxiety, impaired intra-session habituation, evoked circular swimming and disrupted zebrafish shoaling. Additionally, ketamine reduced whole-body cortisol levels and elevated brain c-fos expression in zebrafish. Our findings demonstrate the sensitivity of zebrafish to behavioral and physiological effects of sub-anesthetic doses of ketamine, further supporting the utility of this species as a model for neuropharmacological research, including testing ketamine and related drugs.

Zebrafish (Danio rerio) are becoming increasingly popular in neurobehavioral research. Here, we summarize recent data on behavioral responses of adult zebrafish to a wide spectrum of putative anxiolytic and anxiogenic agents. Using the novel tank test as a sensitive and efficient behavioral assay, zebrafish anxiety-like behavior can be bi-directionally modulated by drugs affecting the gamma-aminobutyric acid, monoaminergic, cholinergic, glutamatergic and opioidergic systems. Complementing human and rodent data, zebrafish drug-evoked phenotypes obtained in this test support this species as a useful model for neurobehavioral and psychopharmacological research.

Animal self‐grooming behavior is becoming increasingly recognized in neurophenotyping research. Rodent grooming and its complex sequencing are sensitive to various genetic and pharmacological manipulations. However, its phenotyping is usually limited to global endpoints such as frequency of bouts and their duration. Here, we used custom‐upgraded HomeCageScan video‐tracking software (Clever Sys. Inc., Reston, VA) to record grooming behavior of adult male C57BL/6J mice in transparent observation cylinders for 5 min. This allowed us not only to perform behavioral quantification of specific grooming patterns (such as paw licking and body/leg grooming) but also analyze their transitions, revealing significant correlations (P<0.0005–0.02, R=0.51–0.70) with manual observations for total number and selected specific grooming transitions. Our data suggests that novel, high‐throughput automated neurophenotyping of grooming behavior can be developed for biomedical research based on this approach.