Abstract
Larval zebrafish (Danio rerio) has the potential to supplement rodent models due to the availability of resource-efficient, high-throughput screening and high-resolution imaging techniques. Although behavioural models are available in larvae, only a few can be employed to assess anxiety. Here we present the swimming plus-maze (SPM) test paradigm, a tool to assess anxiety-related avoidance of shallow water bodies in early developmental stages. The “+” shaped apparatus consists of arms of different depth, representing different levels of aversiveness similarly to the rodent elevated plus-maze. The paradigm was validated (i) in larval and juvenile zebrafish, (ii) after administration of compounds affecting anxiety and (iii) in differentially aversive experimental conditions. Furthermore, we compared the SPM with conventional “anxiety tests” of zebrafish to identify their shared characteristics. We have clarified that the preference of deeper arms is ontogenetically conserved and can be abolished by anxiolytic or enhanced by anxiogenic agents, respectively. The behavioural readout is insensitive to environmental aversiveness and is unrelated to behaviours assessed by conventional tests involving young zebrafish. Taken together, we have developed a sensitive high-throughput test allowing the assessment of anxiety-related responses of zebrafish regardless of developmental stage, granting the opportunity to combine larva-based state-of-the-art methods with detailed behavioral analysis.
Highlights
Mental disorders, those associated with anxiety represent a serious burden on both the individual and the society[1]
We aimed to investigate the nature of connections between each measured variable to draw a general profile for the use of the swimming plus-maze (SPM) test
Our results show that both larval and juvenile zebrafish prefer the deep arms over the shallow arms or the central zone
Summary
Those associated with anxiety represent a serious burden on both the individual and the society[1]. Available behavioural tests utilizing larvae, e.g. the open tank (OT) and the light/dark tank (LDT) test, are based on the natural aversion of exposed[12,13] or poorly lit areas[14,15], but exclude aversion of the water surface, even though it is the most frequently measured, persistently expressed and reproducible defensive response to novelty in adult zebrafish[16,17,18,19,20,21,22,23,24,25,26] This is possibly due to the difficulties with recording the swimming depth of fish in a high-throughput context. To the best of our knowledge, this is the first test that utilizes the anxiety-related avoidance of shallow water in zebrafish regardless of developmental stage, filling an important niche in larva-based high-throughput phenotypic screening
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