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Summary

Abstract

Introduction

Protocol

Representative Results

Discussion

Acknowledgements

Materials

References

Medicine

Boldness, Aggression, and Shoaling Assays for Zebrafish Behavioral Syndromes

Published: August 29th, 2016

DOI:

10.3791/54049

1Department of Biology, Saint Joseph’s University, 2Genomics and Computational Biology, University of Pennsylvania

This manuscript describes the setup, implementation, and analysis of boldness, aggression, and shoaling in zebrafish and testing for the presence of a behavioral syndrome. A standardized approach for behavioral quantification will allow for easier comparison across studies. Modifications to this protocol are possible as each assay can be run individually.

A behavioral syndrome exists when specific behaviors interact under different contexts. Zebrafish have been test subjects in recent studies and it is important to standardize protocols to ensure proper analyses and interpretations. In our previous studies, we have measured boldness by monitoring a series of behaviors (time near surface, latency in transitions, number of transitions, and darts) in a 1.5 L trapezoidal tank. Likewise, we quantified aggression by observing bites, lateral displays, darts, and time near an inclined mirror in a rectangular 19 L tank. By dividing a 76 L tank into thirds, we also examined shoaling preferences. The shoaling assay is a highly customizable assay and can be tailored for specific hypotheses. However, protocols for this assay also must be standardized, yet flexible enough for customization. In previous studies, end chambers were either empty, contained 5 or 10 zebrafish, or 5 pearl danios (D. albolineatus). In the following manuscript, we present a detailed protocol and representative data that accompany successful applications of the protocol, which will allow for replication of behavioral syndrome experiments.

There is a growing body of literature investigating the associations between distinct behaviors within individual animals from a given population. These associations are termed behavioral syndromes, and the measurements typically include boldness, aggression, exploratory behavior, and sociability1-5. Behavioral syndromes are valuable for both direct and indirect reasons. Directly, knowledge of behavioral syndromes can provide a more complete view of evolutionary theory, population structure, and population dynamics3. Indirectly, knowledge regarding behavioral associations may inform fields that quantify behavior such as pharmacology6 ,....

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The following methodologies for the housing, care, and study of zebrafish have been approved by the Saint Joseph's University IACUC.

1. Zebrafish Housing and Care

  1. Obtain the subject zebrafish from a local supplier, or from wild-caught populations. Please note that housing the zebrafish is subject to IACUC guidelines and additional permits are required for housing wild-caught populations.
  2. Identify and separate zebrafish according to sex. Alternatively, skip this step o.......

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Depending on the nature of the study, and specific protocol employed, several distinct results are possible in a behavioral syndromes experiment. The following tables and figures, where indicated, are adapted from our previous study published in the journal Behavioural Processes14 and the journal Zebrafish17. When the proposal (as described above) is carried out in its entirety, two sets of results, 'within assay correlations' and 'between assay correlati.......

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The protocol will determine if there are consistent associations in boldness, aggression, and shoaling behaviors in zebrafish. If there are consistent associations in a given population between any of these behaviors, then a behavioral syndrome is present. By studying a population's natural behavioral syndrome, researchers can have a more complete understanding of its behavioral dynamic, population structure, and possibly evolutionary history3. Furthermore, manipulating the environment that affects these b.......

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This work was supported by a Howard Hughes Medical Institute Education Grant and an internal grant from the Saint Joseph's University chapter of Sigma Xi. We would also like to thank the three anonymous reviewers who helped strengthen the protocol and interpretations.

....

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Name Company Catalog Number Comments
Zebrafish Rack System Aquaneering Inc Cat. # ZS550
Pet Valu Tropical Fish Food, 224.0g Pet Valu Cat. # 31700
Premium Grade Brine Shrimp Eggs, 16 oz Brine Shrimp Direct
1.5 L Trapezoidal Tank Pentair Aquatic Ecosystems Cat. # itsts-a
19L rectangular tank That Fish Place 211932
76L rectangular tank That Fish Place 212180
Hitachi KP-D20A CCD Camera Prescott's, Inc.
Nikon AF Nikkor 35-105mm f/305~4.5s MACRO lens Nikon Corporation
ArtMinds Square Mirror, Value Pack 3"x3" Michaels Cat. # 10334162
Jwatcher
SPSS Statistics Base IBM
R The R Foundation

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