Para-chlorophenylalanine induces multidimensional affective-like behaviors in adult zebrafish

Affective disorders, such as depression and anxiety, pose major challenges to mental health worldwide. Serotonergic dysfunction has been widely implicated in their pathophysiology, although its contribution involves complex and context-dependent mechanisms. To explore the relationship between serotonin depletion and neurobehavioral responses in zebrafish (Danio rerio), animals were treated with para-chlorophenylalanine (pCPA), an inhibitor of tryptophan hydroxylase activity. Fish received two intraperitoneal injections of pCPA (300 mg/kg), with a 24-h interval between injections. Behavioral tests occurred 24 h after the second injection and included the novel tank diving test, tail immobilization, social preference, and shallow water tests. In the novel tank and social preference tests, pCPA-treated fish showed increased geotaxis, with reduced vertical exploration and altered locomotor patterns when facing their conspecifics, suggestive of increased anxiety. Moreover, pCPA exposure increased immobility in both despair-related paradigms, indicative of passive coping in aversive contexts. We also confirmed that pCPA significantly reduced serotonin levels in the brain, showing the effectiveness of the depletion protocol. Overall, these findings expand the behavioral characterization of pCPA-treated adult zebrafish, demonstrating that serotonin depletion modulates multiple behavioral domains. Within a comparative framework, this model provides a platform to investigate multidimensional affective-like phenotypes associated with serotonergic disruption.