1. Ecology
  2. Neuroscience
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Genetic variation in the social environment affects behavioral phenotypes of oxytocin receptor mutants in zebrafish

  1. Diogo Ribeiro
  2. Ana Rita Nunes
  3. Magda Teles
  4. Savani Anbalagan
  5. Janna Blechman
  6. Gil Levkowitz
  7. Rui F Oliveira  Is a corresponding author
  1. Instituto Gulbenkian de Ciência, Portugal
  2. Weizmann Institute of Science, Israel
  3. ReMedy-International Research Agenda Unit, Centre of New Technologies, University of Warsaw, Poland
  4. ISPA – Instituto Universitário, Portugal
  5. Champalimaud Research, Portugal
Short Report
Cite this article as: eLife 2020;9:e56973 doi: 10.7554/eLife.56973
9 figures, 2 tables and 1 additional file

Figures

Genetic variation in the social environment affects zebrafish social behavior.

The contribution of the individual genotype (Gi), the genotype of conspecifics in the social group (Gs) and the interaction between the two (GixGs) to the expression of behavioral phenotypes in zebrafish was assessed by raising oxytocin receptor mutant fish and wild types (focal fish marked with *) in shoals of either mutants or wild types (A). Social preference, measured by the time fish spend near a shoal vs. empty in a choice test (B, upper panel), showed a marginally significant effect of Gs (C; Source data file Figure 1—source data 1). Social habituation, which consisted on a consecutive social preference test exhibited a GixGs effect (D; Source data file Figure 1—source data 2). Social recognition, measured as the discrimination between a novel and a familiar conspecific (E, upper panel), shows a pure G effect (F; Source data file Figure 1—source data 3). Social integration, measured as distance to the centroid of the shoal (G), showed a GixGs effect (H; Source data file Figure 1—source data 4). Social influence, measured by the cohesion of the remaining shoal members (I), also showed a marginally significant GixGs effect (J; Source data file Figure 1—source data 5). Heatmaps show the spatial distribution of a representative oxtr(+/+) individual fish raised in a oxtr(+/+) group, during the entire trial, for both social preference (B, lower panel) and social recognition (E, lower panel). Data is presented as mean ± standard error of the mean (SEM). Sample sizes are nine for heterogeneous groups (i.e. focal individual with different genotype from the remaining individuals in the shoal; mutant focal in WT shoals and WT focal in mutant shoals) and 15 for homogeneous groups (i.e. focal individual with the same genotype of the remaining individuals in the shoal; mutant focal in mutant shoals and WT focal in WT shoals). Different letters indicate significant differences (p<0.05) between treatments as assessed by Tukey post-hoc tests following a two-way ANOVA (D,H,J; see Table 1). An asterisk indicates a Gi main effect in F.

Figure 1—source data 1

Effects of individual and conspecifics genotype on Social Preference.

https://cdn.elifesciences.org/articles/56973/elife-56973-fig1-data1-v2.xlsx
Figure 1—source data 2

Effects of individual and conspecifics genotype on social habituation.

https://cdn.elifesciences.org/articles/56973/elife-56973-fig1-data2-v2.xlsx
Figure 1—source data 3

Effects of individual and conspecifics genotype on social recognition.

https://cdn.elifesciences.org/articles/56973/elife-56973-fig1-data3-v2.xlsx
Figure 1—source data 4

Effects of individual and conspecifics genotype on social integration.

https://cdn.elifesciences.org/articles/56973/elife-56973-fig1-data4-v2.xlsx
Figure 1—source data 5

Effects of individual and conspecifics genotype on social influence.

https://cdn.elifesciences.org/articles/56973/elife-56973-fig1-data5-v2.xlsx
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Tables

Table 1
Effect of genotype of the focal individual (Gi), genotype of conspecifics present in its social environment (Gs) and the interaction between the two (GixGs) on zebrafish social behavior was assessed using a two-way ANOVA.

~ indicates marginally significant, *p<0.05, **p<0.01, ***p<0.001. (Source data files Figure 1—source datas 15).

Social preference
d.f.Mean squaresFSignificancePartial η2
Gi10.0231.7310.1950.038
Gs10.0503.7880.058~0.079
Gi x Gs10.0010.0490.8250.001
Error440.013
Habituation
d.f.Mean squaresFSignificancePartial η2
Gi10.05813.9270.001 **0.240
Gs10.0081.9360.1710.042
Gi x Gs10.0245.6420.022 *0.114
Error440.004
Social recognition
d.f.Mean squaresFSignificancePartial η2
Gi10.2137.6000.008 **0.147
Gs10.0050.1890.6660.004
Gi x Gs10.0010.0410.8410.001
Error440.028
Social group integration
d.f.Mean squaresFSignificancePartial η2
Gi139.48624.370<0.001 ***0.356
Gs112.5657.7550.008 **0.150
Gi x Gs112.8117.9070.007 **0.152
Error441.620
Social group dispersion
d.f.Mean squaresFSignificancePartial η2
Gi1174.3664.3090.044 *0.089
Gs1657.22116.240<0.001 ***0.270
Gi x Gs1122.9803.0390.0880.065
Error4440.469
Key resources table
Reagent type
(species) or resource
DesignationSource or referenceIdentifiersAdditional
information
Genetic reagent, TL (Danio rerio)oxtr mutant lineNunes et al., 2020ZDB-ALT-190830–1
Commercial assay or kitNucleoSpin TissueMACHEREY-NAGEL# 740952.50For oxtr mutant genotyping
Sequence-based reagentsense 5’-TGCGCGAGGAAAACTAGTT-3’SigmaFor oxtr mutant genotyping
Sequence-based reagentantisense 5’-AGCAGACACTCAGAATGGTCA-3’SigmaFor oxtr mutant genotyping
Software, , algorithmSPSS 25.0SPSSRRID:SCR_002865
Software, , algorithmImagej (Fiji)Schindelin et al., 2012RRID:SCR_003070
Software, , algorithmEthovision XT 11.5Noldus Technologywww.noldus.com/ethovision
Software, , algorithmGraphPad Prism version 6.0 cGraphPad software, San Diego, California, USAwww.graphpad.com
OtherB and W mini surveillance cameraHenelec 300BAcquisition rate of 30 fps
OtherWebcamerasLogitech HD C525Acquisition rate of 30 fps

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