Acute effects of ethanol on behavioral responses of male and female zebrafish in the open field test with the influence of a non-familiar object.

In this report, we investigate whether the acute effects of different ethanol (EtOH) concentrations are sex-dependent in zebrafish subjected to the open field test (OFT) with the influence of a non-familiar object. Male and female zebrafish were separated into four groups and exposed to EtOH (0%, 0.25%, 0.5%, or 1.0% v/v) for 1 h. Fish were tested individually in the OFT and the tank was divided into three areas: periphery, intermediate, and center area. An object (black sphere; diameter: 1 cm) was placed in the center of the tank and behaviors were recorded for 5 min. At the baseline, females had a distinct exploratory activity and interaction pattern with the object, reflecting a more anxious and shyer behavior in relation to males. Females exposed to 0.5% EtOH performed more rapid investigation to the object than males, while 1.0% EtOH reduced locomotion in both sexes and increased immobility only in males. Principal component analyses revealed that anxiety-like behaviors, exploratory activity, and locomotion were the components that most accounted for total variances. Collectively, our novel findings show the existence of a sex-dependent effect in the zebrafish models acutely exposed to EtOH tested in the OFT with a non-familiar object.