Author:
Ferreira Jorge M.,Silva Joana,Barros Sofia,Caetano Inês,Fernandes Pedro,Olsson Anna,Valentim Ana M.
Abstract
AbstractOpen-source validated tools are important for affordable high-quality science. Zebrafish model is often used in behavioral sciences, thus validated tools must be developed for this species. We aimed to implement the memory task one-trial inhibitory avoidance test in our laboratory by creating a custom-built 3D printed apparatus and custom-made hardware and software using microcontrollers; this decreases the costs and increases methodological flexibility. In this task, a mild electric shock (3.3 ± 0.3V and 2A for 5sec) is used as an aversive stimulus for the one-trial inhibitory avoidance task based on classical conditioning. For this study we used 72-adult zebrafish from the AB and TU strain. The aversive stimulus caused a robust and long-lasting memory that was learned in one session. The apparatus consisted in one aquarium with two compartments (white and black) where, during the conditioning session, the shock was delivered in the preferred side (black), followed by 30min of immersion in 20μM MK-801, a compound known for inducing amnesia (amnesic group, n= 20 for AB and 17 for TU), or in clean water (control, n= 20 for AB and 15 for TU). Both the AB and TU strain animals from the control group took more time to enter in the black compartment after conditioning (p≤0.039), indicating avoidance for the black side, while the latency was not altered in the MK-801 group, demonstrating impaired memory. In summary, this open-source apparatus is an affordable and validated option for memory assessment for the two most used strains of zebrafish in research.
Publisher
Cold Spring Harbor Laboratory
Reference32 articles.
1. E.E. Commission , report on the statistics on the use of animals for scientific purposes in the Member States of the European Union in 2015-2017, Brussels 5 (2020).
2. C.M. Loss , F.F. Melleu , K. Domingues , C. Lino-de-Oliveira , G.G. Viola , Combining Animal Welfare With Experimental Rigor to Improve Reproducibility in Behavioral Neuroscience, Front Behav Neurosci 15 (2021).
3. W.M.S. Russell , R.L. Burch , The principles of humane experimental technique, Methuen 1959.
4. J.M. Ferreira , I.A.S. Olsson , A.M. Valentim , Disentangling the terminology of the light/dark and white/black test in zebrafish, (2023).
5. J.D. Sweatt , Rodent behavioral learning and memory models, Mechanisms of Memory, 2nd ed. Elsevier, UK (2010) 77–103.