Effects of diazepam and nortriptyline on behavioral changes and chronic stress-induced depression in zebrafish (Danio rerio)

Document Type : Research Paper

Authors

1 Doctor of Veterinary Medicine, Babol Branch, Islamic Azad University, Babol, Iran

2 Assistant Professor in Department of Veterinary, Babol Branch, Islamic Azad University, Babol, Iran

3 Associate Professor in Department of Physiology, Mazandaran University of Medical Science, Sari, Iran

10.22124/japb.2025.30222.1566

Abstract

Depression is a mental disorder that according to global statistics, is becoming increasingly prevalent, highlighting the urgent need for further research on depression models and the development of novel antidepressant drugs. This study aimed to evaluate the individual and combined effects of diazepam and nortriptyline on behavioral and biochemical parameters in zebrafish (Danio rerio). A total of 384 zebrafish were initially divided into two main groups: a control group (unstressed) and a UCS group (subjected to stress) and within each group, fish were assigned to one of four subgroups: control, diazepam, nortriptyline, and diazepam + nortriptyline (eight treatments with three replicates). To induce depression-like states in zebrafish, the Unpredictable Chronic Stress (UCS) protocol was applied. Behavioral assessments were conducted using the Novel Tank Test (NTT), and biochemical analysis was carried out by measuring whole-body cortisol levels. Diazepam and nortriptyline led to significant improvements in behavioral parameters, including a notable increase in time spent in the upper zone and number of transitions to the upper zone, and a significant reduction in total distance traveled. These behavioral improvements were accompanied by a marked decrease in cortisol levels (from 1.01±0.07 to 0.38±0.16ng/mL). The results are consistent with previous studies, confirming that the UCS protocol induces behavioral and neuroendocrine changes in zebrafish, which can be attenuated by the administration of diazepam and nortriptyline. Moreover, this study reinforces the utility of zebrafish as a model for investigating the behavioral and physiological impacts of stress.

Keywords

Main Subjects

References

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Aquatic Physiology and Biotechnology
Volume 13, Issue 4
March 2026
Pages 117-137

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