The role of CRH in behavioral responses to acute restraint stress in zebrafish

In teleosts, changes in swimming, exploring, general locomotor activity, and anxious state can be a response to stress mediated by the corticotropin-releasing hormone system activation and its effects on glucocorticoid levels. Zebrafish has been widely used to study neuropharmacology and has become a promising animal model to investigate neurobehavioral mechanisms of stress. In this report the animals were submitted to acute restraint stress for different time lengths (15, 60 and 90 min) for further evaluation of behavioral patterns, whole-body cortisol content, and corticotropin-releasing hormone expression. The results demonstrated an increase in the locomotor activity and an alteration in the swimming pattern during a 5-min trial after the acute restraint stress. Interestingly, all groups of fish tested in the novel tank test exhibited signs of anxiety as evaluated by the time spent in the bottom of the tank. Whole-body cortisol content showed a positive correlation with increased behavioral indices of locomotion in zebrafish whereas molecular analysis of corticotropin-releasing hormone showed a late reduction of mRNA expression (90 min). Altogether, we present a model of acute restraint stress in zebrafish, confirmed by elevated cortisol content, as a valid and reliable model to study the biochemical basis of stress behavior, which seems to be accompanied by a negative feedback of corticotropin-release hormone mRNA expression.     

The use of the zebrafish model in stress research

The study of the causes and mechanisms underlying psychiatric disorders requires the use of non-human models for the test of scientific hypotheses as well as for use in pre-clinical drug screening and discovery. This review argues in favor of the use of zebrafish as a novel animal model to study the impact of early (stressful) experiences on the development of differential stress phenotypes in later life. This phenomenon is evolutionary conserved among several vertebrate species and has relevance to the etiology of psychiatric disorders. Why do we need novel animal models? Although significant progress has been achieved with the use of traditional mammalian models, there are major pitfalls associated with their use that impedes progress on two major fronts: 1) uncovering of the molecular mechanisms underlying aspects of compromised (stress-exposed) brain development relevant to the etiology of psychiatric disorders, and 2) ability to develop high-throughput technology for drug discovery in the field of psychiatry. The zebrafish model helps resolve these issues.Here we present a conceptual framework for the use of zebrafish in stress research and psychiatry by addressing three specific domains of application: 1) stress research, 2) human disease mechanisms, and 3) drug discovery. We also present novel methodologies associated with the development of the zebrafish stress model and discuss how such methodologies can contribute to remove the main bottleneck in the field of drug discovery.     

Behavioral changes induced by long-term proline exposure are reversed by antipsychotics in zebrafish

Hyperprolinemia is an inherited disorder of proline metabolism and patients affected by this disease may present neurological manifestations, including seizures and cognitive dysfunctions. Moreover, an association between adulthood schizoaffective disorders and moderate hyperprolinemia has been reported. However, the mechanisms underlying these behavioral phenotypes still remain unclear. In the present study, we investigated the effect of proline treatments on behavioral parameters in zebrafish, such as locomotor activity, anxiety, and social interaction. Adult zebrafish (Danio rerio) were exposed to proline (1.5 and 3.0 mM) during 1h or 7 days (short- or long-term treatments, respectively). Short-term proline exposure did not promote significant changes on the behavioral parameters observed. Long-term exposure at 1.5 mM proline significantly increased the number of line crossing (47%), the total distance (29%), and the mean speed (33%) when compared to control group. A significant increase in the time spent in the upper portion of the test tank was also observed after this treatment (91%), which may be interpreted as an indicator of anxiolytic behavior. Proline at 1.5 mM also induced social interaction impairment (78%), when compared to the untreated group after long-term treatment. Moreover, these proline-induced behavioral changes in zebrafish were completely reversed by acute administration of an atypical antipsychotic drug (sulpiride), but not by a typical (haloperidol). These findings demonstrate that proline is able to induce schizophrenia-like symptoms in zebrafish, which reinforce the use of this species as a complementary vertebrate model for studying behavioral phenotypes associated with neurological dysfunctions characteristic of metabolic diseases.