Comparative Analyses of Zebrafish Anxiety-Like Behavior Using Conflict-Based Novelty Tests

Abstract

Modeling of stress and anxiety in adult zebrafish (Danio rerio) is increasingly utilized in neuroscience research and central nervous system (CNS) drug discovery. Representing the most commonly used zebrafish anxiety models, the novel tank test (NTT) focuses on zebrafish diving in response to potentially threatening stimuli, whereas the light-dark test (LDT) is based on fish scototaxis (innate preference for dark vs. bright areas). Here, we systematically evaluate the utility of these two tests, combining meta-analyses of published literature with comparative in vivo behavioral and whole-body endocrine (cortisol) testing. Overall, the NTT and LDT behaviors demonstrate a generally good cross-test correlation in vivo, whereas meta-analyses of published literature show that both tests have similar sensitivity to zebrafish anxiety-like states. Finally, NTT evokes higher levels of cortisol, likely representing a more stressful procedure than LDT. Collectively, our study reappraises NTT and LDT for studying anxiety-like states in zebrafish, and emphasizes their developing utility for neurobehavioral research. These findings can help optimize drug screening procedures by choosing more appropriate models for testing anxiolytic or anxiogenic drugs.

Response mechanisms to joint exposure of triclosan and its chlorinated derivatives on zebrafish (Danio rerio) behavior

“…Previous studies reported a variety of behavioral tests and models such as the open field test, shoaling test, and impoverished housing, among which the light-to-dark test, CPP, T-maze and BDT were the most popular experimental models for zebrafish behavioral studies (Stewart et al, 2012) (Kysil et al, 2017); investigated the comparative analyses of zebrafish anxiety-like behavior using a novel tank test (NTT) and light-dark test (LTD), and found that some NTT and CPP endpoints, and endocrine (cortisol) levels were closely related to anxiety. Our results are consistent with these investigations demonstrating that CPP, BDT, ERa and ERb were linked with anxiety-or autism-like diseases.…”

The Integration of Sociality, Monoamines, and Stress Neuroendocrinology in Fish Models: Applications in the Neurosciences

“…Two assays -the novel tank test (Egan et al, 2009) and the light/dark test (Maximino et al, 2010b) -are widely used in this regard, with good pharmacological validation and construct validity for both tests Kysil et al, 2017); in both cases, threat is merely potential, as no predator or partial predator stimuli are present. In the novel tank test, the novelty of the environment elicit bottom-dwelling (also termed a "diving" response) that is associated with erratic swimming and freezing, representing defensive behaviour in this assay (Egan et al, 2009).…”

“…In the light/dark test, the preference for a black compartment over a white one, associated with risk assessment behaviours, erratic swimming, thigmotaxis, and freezing is interpreted as defensive behaviour (Maximino et al, 2010b). Moreover, a relationship between stress and behaviour in these tests is established, as both acute (Giacomini et al, 2016) and chronic (Chakravarty et al, 2013;Marcon et al, 2016) stress increases defensive behaviour in these assays, and exposure to the apparatuses induce cortisol levels (Kysil et al, 2017). A causal link between neuroendocrine and behavioural responses to stress has not been ascertained; since these are modulated by monoamines, these neurotransmitters could link both functions.…”

The Integration of Sociality, Monoamines, and Stress Neuroendocrinology in Fish Models: Applications in the Neurosciences

“…Nonetheless, zebrafish is the most widely studied species when it comes to fear, anxiety, and stress (Jesuthasan & Mathuru, 2008;Maximino et al, 2010a;Kalueff et al, 2012;Gerlai, 2013). Two assays -the novel tank test (Egan et al, 2009) and the light/dark test (Maximino et al, 2010b) -are widely used in this species, with good pharmacological validation and construct validity for both tests Kysil et al, 2017). In the novel tank test, the novelty of the environment elicit bottom-dwelling (also termed a "diving" response) that is associated with erratic swimming and freezing, representing anxiety-like behavior in this assay (Egan et al, 2009).…”

“…Moreover, a relationship between stress and behaviour in these tests is established, as both acute (Giacomini et al, 2016) and chronic (Chakravarty et al, 2013;Marcon et al, 2016) stress increases anxiety-like behaviour in these assays, and exposure to the apparatuses induce cortisol levels (Kysil et al, 2017). A causal link between neuroendocrine and behavioural responses to stress has not been ascertained; since these are modulated by monoamines, these upstream modulators could link both functions.…”