The Visually Mediated Social Preference Test: A Novel Technique to Measure Social Behavior and Behavioral Disturbances in Zebrafish

  • William H. J. Norton
  • Line Manceau
  • Florian ReichmannEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 2011)


Zebrafish are an emerging model in behavioral neuroscience. They display a wide range of measurable behaviors such as locomotion, aggression, anxiety, learning and memory, and social behavior. In addition, the relative ease of genetic manipulation and the increasing availability of disease models mean that zebrafish have gained in popularity as an animal model for various neurological and psychiatric diseases including autism spectrum disorder (ASD). In order to better characterize social behavior and behavioral abnormalities in zebrafish, we have developed the visually mediated social preference (VMSP) test, a novel assay to measure social preference and social novelty in two consecutive 5-min sessions. Using recording and video tracking, the time spent in different areas of the tank, the time spent immobile, swimming speed, and distance moved can be easily measured and analyzed. Untreated experimentally naive AB WT zebrafish typically show a strong preference for spending time near and interacting with a compartment containing unfamiliar conspecifics over the empty compartments during session 1 and a stronger preference for a group of unfamiliar zebrafish over familiar conspecifics from session 1, during session 2 of the test. Research in our lab has shown that the VMSP is suitable to measure the social behavior of individual zebrafish, to uncover social phenotypes of mutant strains, and to better understand animal models of disease that include impaired sociability such as ASD. The current paper provides a step-by-step guide on how to implement and perform this test and highlights important considerations for data acquisition, analysis, and interpretation.

Key words

Social behavior Social interaction Social preference Social novelty Zebrafish Autism spectrum disorder 



We thank Hector Carreno-Gutierrez, Elisa Dalla Vecchia, and Ceinwen Tilley for the critical reading and comments on the manuscript and Carl Breaker for technical assistance. This work was supported by the Austrian Science Fund (grant number J4090-B29).


  1. 1.
    Stewart AM, Grieco F, Tegelenbosch RA, Kyzar EJ, Nguyen M, Kaluyeva A, Song C, Noldus LP, Kalueff AV (2015) A novel 3D method of locomotor analysis in adult zebrafish: implications for automated detection of CNS drug-evoked phenotypes. JNeurosciMethods 255:66–74Google Scholar
  2. 2.
    Jones LJ, Norton WH (2015) Using zebrafish to uncover the genetic and neural basis of aggression, a frequent comorbid symptom of psychiatric disorders. Behav Brain Res 276:171–180CrossRefGoogle Scholar
  3. 3.
    Maximino C, de Brito TM, da Silva Batista AW, Herculano AM, Morato S, Gouveia A Jr (2010) Measuring anxiety in zebrafish: a critical review. Behav Brain Res 214:157–171CrossRefGoogle Scholar
  4. 4.
    Bailey JM, Oliveri AN, Levin ED (2015) Pharmacological analyses of learning and memory in zebrafish (Danio rerio). Pharmacol Biochem Behav 139(Pt B):103–111CrossRefGoogle Scholar
  5. 5.
    Miller N, Gerlai R (2012) From schooling to shoaling: patterns of collective motion in zebrafish (Danio rerio). PLoS One 7:e48865CrossRefGoogle Scholar
  6. 6.
    Norton WH (2013) Toward developmental models of psychiatric disorders in zebrafish. Front Neural Circuits 7:79CrossRefGoogle Scholar
  7. 7.
    Kalueff AV, Stewart AM, Gerlai R (2014) Zebrafish as an emerging model for studying complex brain disorders. Trends Pharmacol Sci 35:63–75CrossRefGoogle Scholar
  8. 8.
    Shams S, Rihel J, Ortiz JG, Gerlai R (2018) The zebrafish as a promising tool for modeling human brain disorders: a review based upon an IBNS Symposium. Neurosci Biobehav Rev 85:176–190CrossRefGoogle Scholar
  9. 9.
    Meshalkina DA, N Kizlyk M, V Kysil E, Collier AD, Echevarria DJ, Abreu MS, Barcellos LJG, Song C, Warnick JE, Kyzar EJ, Kalueff AV (2018) Zebrafish models of autism spectrum disorder. Exp Neurol 299:207–216CrossRefGoogle Scholar
  10. 10.
    Huang J, Zhong Z, Wang M, Chen X, Tan Y, Zhang S, He W, He X, Huang G, Lu H, Wu P, Che Y, Yan YL, Postlethwait JH, Chen W, Wang H (2015) Circadian modulation of dopamine levels and dopaminergic neuron development contributes to attention deficiency and hyperactive behavior. J Neurosci 35:2572–2587CrossRefGoogle Scholar
  11. 11.
    Grone BP, Baraban SC (2015) Animal models in epilepsy research: legacies and new directions. Nat Neurosci 18:339–343CrossRefGoogle Scholar
  12. 12.
    Newman M, Ebrahimie E, Lardelli M (2014) Using the zebrafish model for Alzheimer’s disease research. Front Genet 5:189PubMedPubMedCentralGoogle Scholar
  13. 13.
    Das S, Rajanikant GK (2014) Huntington disease: can a zebrafish trail leave more than a ripple? Neurosci Biobehav Rev 45:258–261CrossRefGoogle Scholar
  14. 14.
    Fonseka TM, Wen XY, Foster JA, Kennedy SH (2016) Zebrafish models of major depressive disorders. J Neurosci Res 94:3–14CrossRefGoogle Scholar
  15. 15.
    Matsui H, Takahashi R (2018) Parkinson’s disease pathogenesis from the viewpoint of small fish models. J Neural Transm (Vienna) 125:25–33CrossRefGoogle Scholar
  16. 16.
    Miller N, Gerlai R (2007) Quantification of shoaling behaviour in zebrafish (Danio rerio). Behav Brain Res 184:157–166CrossRefGoogle Scholar
  17. 17.
    Perez-Escudero A, Vicente-Page J, Hinz RC, Arganda S, de Polavieja GG (2014) idTracker: tracking individuals in a group by automatic identification of unmarked animals. Nat Methods 11:743–748CrossRefGoogle Scholar
  18. 18.
    Toth I, Neumann ID (2013) Animal models of social avoidance and social fear. Cell Tissue Res 354:107–118CrossRefGoogle Scholar
  19. 19.
    Nadler JJ, Moy SS, Dold G, Trang D, Simmons N, Perez A, Young NB, Barbaro RP, Piven J, Magnuson TR, Crawley JN (2004) Automated apparatus for quantitation of social approach behaviors in mice. Genes Brain Behav 3:303–314CrossRefGoogle Scholar
  20. 20.
    Gerlach G, Hodgins-Davis A, Avolio C, Schunter C (2008) Kin recognition in zebrafish: a 24-hour window for olfactory imprinting. Proc Biol Sci 275:2165–2170CrossRefGoogle Scholar
  21. 21.
    Parichy DM, Elizondo MR, Mills MG, Gordon TN, Engeszer RE (2009) Normal table of postembryonic zebrafish development: staging by externally visible anatomy of the living fish. Dev Dyn 238:2975–3015CrossRefGoogle Scholar
  22. 22.
    Vergauwen L, Knapen D, Hagenaars A, Boeck GD, Blust R (2013) Assessing the impact of thermal acclimation on physiological condition in the zebrafish model. J Comp Physiol B 183:109–121CrossRefGoogle Scholar
  23. 23.
    Collymore C, Tolwani RJ, Rasmussen S (2015) The behavioral effects of single housing and environmental enrichment on adult zebrafish (Danio rerio). J Am Assoc Lab Anim Sci 54:280–285PubMedPubMedCentralGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • William H. J. Norton
    • 1
  • Line Manceau
    • 1
  • Florian Reichmann
    • 2
    Email author
  1. 1.Department of Neuroscience, Psychology and BehaviourUniversity of LeicesterLeicesterUK
  2. 2.Otto Loewi Research CentreMedical University of GrazGrazAustria

Personalised recommendations