Abstract
Apart from morphological, biochemical, and genetic alterations induced by teratogen compounds, there is an increased interest in characterizing behavioral alterations. Behavior is a sensitive parameter that can provide information regarding developmental disruptions non-invasively. Behavioral disturbances interfere with animals’ capacity to cope with the environment, having an impact on the organism’s life. Hereby, behavioral assays consisting of recording larvae in multi-well plates, Petri dishes, or cuvettes and video analysis using adequate software, allowing teratogen screening of behavior, are proposed. Examples of how to evaluate locomotor, anxiety-like and avoidance-like behaviors, and the integrity of sensory-motor functions and learning are discussed in this chapter.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
He J-H, Gao J-M, Huang C-J et al (2014) Zebrafish models for assessing developmental and reproductive toxicity. Neurotoxicol Teratol 42(Supplement C):35–42. https://doi.org/10.1016/j.ntt.2014.01.006
Cottrell JE, Hartung J (2012) Developmental disability in the young and postoperative cognitive dysfunction in the elderly after anesthesia and surgery: do data justify changing clinical practice? Mt Sinai J Med 79(1):75–94. https://doi.org/10.1002/msj.21283
Felix LM, Serafim C, Valentim AM et al (2016) Embryonic stage-dependent teratogenicity of ketamine in zebrafish (Danio rerio). Chem Res Toxicol 29(8):1298–1309. https://doi.org/10.1021/acs.chemrestox.6b00122
Félix L, Coimbra AM, Valentim AM et al (2019) Review on the use of zebrafish embryos to study the effects of anesthetics during early development. Crit Rev Toxicol:1–14. https://doi.org/10.1080/10408444.2019.1617236
Sharma S, Coombs S, Patton P et al (2009) The function of wall-following behaviors in the Mexican blind cavefish and a sighted relative, the Mexican tetra (Astyanax). J Comp Physiol A Neuroethol Sens Neural Behav Physiol 195(3):225–240. https://doi.org/10.1007/s00359-008-0400-9
Treit D, Fundytus M (1988) Thigmotaxis as a test for anxiolytic activity in rats. Pharmacol Biochem Behav 31(4):959–962
Schnorr SJ, Steenbergen PJ, Richardson MK et al (2012) Measuring thigmotaxis in larval zebrafish. Behav Brain Res 228(2):367–374. https://doi.org/10.1016/j.bbr.2011.12.016
Fontana BD, Parker MO (2022) The larval diving response (LDR): validation of an automated, high-throughput, ecologically relevant measure of anxiety-related behavior in larval zebrafish (Danio rerio). J Neurosci Methods 381:109706. https://doi.org/10.1016/j.jneumeth.2022.109706
Bishop BH, Spence-Chorman N, Gahtan E (2016) Three-dimensional motion tracking reveals a diving component to visual and auditory escape swims in zebrafish larvae. J Exp Biol 219(Pt 24):3981–3987. https://doi.org/10.1242/jeb.147124
Grillon C (2008) Models and mechanisms of anxiety: evidence from startle studies. Psychopharmacology 199(3):421–437. https://doi.org/10.1007/s00213-007-1019-1
Roberts AC, Reichl J, Song MY et al (2011) Habituation of the C-start response in larval zebrafish exhibits several distinct phases and sensitivity to NMDA receptor blockade. PLoS One 6(12):e29132. https://doi.org/10.1371/journal.pone.0029132
Sztal TE, Ruparelia AA, Williams C et al (2016) Using touch-evoked response and locomotion assays to assess muscle performance and function in zebrafish. J Vis Exp 116. https://doi.org/10.3791/54431
Creton R (2009) Automated analysis of behavior in zebrafish larvae. Behav Brain Res 203(1):127–136. https://doi.org/10.1016/j.bbr.2009.04.030
Felix LM, Antunes LM, Coimbra AM et al (2017) Behavioral alterations of zebrafish larvae after early embryonic exposure to ketamine. Psychopharmacology 234(4):549–558. https://doi.org/10.1007/s00213-016-4491-7
Egan RJ, Bergner CL, Hart PC et al (2009) Understanding behavioral and physiological phenotypes of stress and anxiety in zebrafish. Behav Brain Res 205(1):38–44. https://doi.org/10.1016/j.bbr.2009.06.022
Buske C, Gerlai R (2014) Diving deeper into zebrafish development of social behavior: analyzing high resolution data. J Neurosci Methods 234:66–72. https://doi.org/10.1016/j.jneumeth.2014.06.019
Kalueff AV, Gebhardt M, Stewart AM et al (2013) Towards a comprehensive catalog of zebrafish behavior 1.0 and beyond. Zebrafish 10(1):70–86. https://doi.org/10.1089/zeb.2012.0861
Ali S, Champagne DL, Alia A et al (2011) Large-scale analysis of acute ethanol exposure in zebrafish development: a critical time window and resilience. PLoS One 6(5):e20037. https://doi.org/10.1371/journal.pone.0020037
Norton WHJ (2012) Measuring larval zebrafish behavior: locomotion, thigmotaxis, and startle. In: Kalueff AV, Stewart AM (eds) Zebrafish protocols for neurobehavioral research. Humana Press, Totowa, pp 3–20. https://doi.org/10.1007/978-1-61779-597-8_1
Best JD, Berghmans S, Hunt JJFG et al (2007) Non-associative learning in larval zebrafish. Neuropsychopharmacology 33(5):1206–1215. http://www.nature.com/npp/journal/v33/n5/suppinfo/1301489s1.html
Clark DT (1981) Visual responses in developing zebrafish (Brachydanio Rerio). University of Oregon, Eugene
Emran F, Rihel J, Adolph AR et al (2007) OFF ganglion cells cannot drive the optokinetic reflex in zebrafish. Proc Natl Acad Sci U S A 104(48):19126–19131. https://doi.org/10.1073/pnas.0709337104
Strahle U, Scholz S, Geisler R et al (2012) Zebrafish embryos as an alternative to animal experiments-A commentary on the definition of the onset of protected life stages in animal welfare regulations. Reprod Toxicol 33(2):128–132. https://doi.org/10.1016/j.reprotox.2011.06.121
Smith LL, Beggs AH, Gupta VA (2013) Analysis of skeletal muscle defects in larval zebrafish by birefringence and touch-evoke escape response assays. J Vis Exp 82:e50925. https://doi.org/10.3791/50925
Acknowledgments
The author would like to thank Margarida Monteiro (M.S.), Jorge Ferreira (M.S., Instituto de Investigação e Inovação em Saúde, Porto, Portugal), Luís Félix (PhD, Universidade de Trás-os-Montes e Alto Douro, Vila Real, Portugal), Isabel Silveira (PhD) and Joana Loureiro (PhD), both from the Instituto de Investigação e Inovação em Saúde, Porto, Portugal, for their support in the behavioral tests and analysis. The author also would like to thank Pedro Silva for his support in the Arduino assembly and programming. The first edition of this work was supported by a postdoctoral fellowship SFRH/BPD/103006/2014 issued by Fundação para a Ciência e Tecnologia (FCT), Portugal.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2024 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Valentim, A.M. (2024). Behavioral Profiling of Zebrafish (Danio rerio) Larvae: Activity, Anxiety, Avoidance, and Startle Response. In: Félix, L. (eds) Teratogenicity Testing. Methods in Molecular Biology, vol 2753. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3625-1_26
Download citation
DOI: https://doi.org/10.1007/978-1-0716-3625-1_26
Published:
Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-3624-4
Online ISBN: 978-1-0716-3625-1
eBook Packages: Springer Protocols