Abstract
RationaleTriangulation of approaches (i.e., using several tests of the same construct) can be extremely useful for increasing the robustness of the findings being widely used when working with behavioral testing, especially when using rodents as a translational model. Although zebrafish are widely used in neuropharmacology research due to their high-throughput screening potential for new therapeutic drugs, behavioral test battery effects following pharmacological manipulations are still unknown.MethodsHere, we tested the effects of an anxiety test battery and test time following pharmacological manipulations in zebrafish by using two behavioral tasks: the novel tank diving task (NTT) and the light–dark test (LDT). Fluoxetine and conspecific alarm substance (CAS) were chosen to induce anxiolytic and anxiogenic-like behavior, respectively.ResultsFor non-drug-treated animals, no differences were observed for testing order (NTT → LDT or LDT → NTT) and there was a strong correlation between performances on the two behavioral tasks. However, we found that during drug treatment, NTT/LDT responses are affected by the tested order depending on the test time being fluoxetine effects higher at the second behavioral task (6 min later) and CAS effects lower across time.ConclusionsOverall, our data supports the use of baseline behavior assessment using this anxiety test battery. However, when working with drug exposure, data analysis must carefully consider time-drug-response and data variability across behavioral tasks.
Highlights
In behavioral research, triangulation of approaches can be extremely useful for increasing the robustness of the findings, and increase the confidence in the validity of the results (Stegenga 2009)
Two-way ANOVA with multiple testing and substance exposure as fixed factors were used to compare anxiety endpoints (NTT, time spent in top of the tank and distance traveled; light–dark test (LDT), time in the dark area)
Fluoxetine anxiolytic‐like effect is increased with a time delay or when secondly tested In Fig. 3, the behavioral phenotype of fish exposed to fluoxetine compared to controls tested in the novel tank diving task (NTT) and LDT is compared
Summary
Triangulation of approaches (i.e., using several tests of the same construct) can be extremely useful for increasing the robustness of the findings, and increase the confidence in the validity of the results (Stegenga 2009). Behavioral test batteries are widely employed, where animals are tested in multiple behavioral tasks either on the same day or across weeks, and the results are triangulated to gain a more robust operational definition of the target behavior (Paylor et al 2006). While behavioral test batteries are common in rodent research, they have been less widely employed in zebrafish behavioral studies. The systematic assessment of the impact of multiple behavioral tests on zebrafish performance in the assays is yet to be carried out
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