Behavior Genetics

, Volume 47, Issue 1, pp 88–101 | Cite as

Genetics of Interactive Behavior in Silver Foxes (Vulpes vulpes)

  • Ronald M. Nelson
  • Svetlana V. Temnykh
  • Jennifer L. Johnson
  • Anastasiya V. Kharlamova
  • Anastasiya V. Vladimirova
  • Rimma G. Gulevich
  • Darya V. Shepeleva
  • Irina N. Oskina
  • Gregory M. Acland
  • Lars Rönnegård
  • Lyudmila N. Trut
  • Örjan Carlborg
  • Anna V. Kukekova
Original Research


Individuals involved in a social interaction exhibit different behavioral traits that, in combination, form the individual’s behavioral responses. Selectively bred strains of silver foxes (Vulpes vulpes) demonstrate markedly different behaviors in their response to humans. To identify the genetic basis of these behavioral differences we constructed a large F2 population including 537 individuals by cross-breeding tame and aggressive fox strains. 98 fox behavioral traits were recorded during social interaction with a human experimenter in a standard four-step test. Patterns of fox behaviors during the test were evaluated using principal component (PC) analysis. Genetic mapping identified eight unique significant and suggestive QTL. Mapping results for the PC phenotypes from different test steps showed little overlap suggesting that different QTL are involved in regulation of behaviors exhibited in different behavioral contexts. Many individual behavioral traits mapped to the same genomic regions as PC phenotypes. This provides additional information about specific behaviors regulated by these loci. Further, three pairs of epistatic loci were also identified for PC phenotypes suggesting more complex genetic architecture of the behavioral differences between the two strains than what has previously been observed.


Behavior genetics Social behavior Quantitative trait loci Domestication Aggression Epistasis Vulpes vulpes Canis familiaris 



We are grateful to Irina V. Pivovarova, Tatyana I. Semenova, and all the animal keepers at the ICG experimental farm for research assistance. We thank K. Gordon Lark and Kevin Chase for advice and important discussions. The project was supported by National Institutes of Health Grant MH077811, NIH FIRCA Grant TW008098, USDA Federal Hatch Project #538922, Program of the Siberian Branch of the Russian Academy of Sciences #0324-2015-0007, Grant #13-04-00420 from the Russian Fund for Basic Research, and Campus Research Board Grant from the University of Illinois at Urbana-Champaign.

Conflict of interest

Ronald M. Nelson, Svetlana V. Temnykh, Jennifer L. Johnson, Anastasiya V. Kharlamova, Anastasiya V. Vladimirova, Rimma G. Gulevich, Darya V. Shepeleva, Irina N. Oskina, Gregory M. Acland, Lars Rönnegård, Lyudmila N. Trut, Örjan Carlborg, Anna V. Kukekova declare that they have no conflict of interest.

Human and animal rights and informed consent

All institutional and national guidelines for the care and use of laboratory animals were followed. All animal procedures at the Institute of Cytology and Genetics of the Russian Academy of Sciences complied with standards for humane care and use of laboratory animals by foreign institutions.

Supplementary material

10519_2016_9815_MOESM1_ESM.xlsx (51 kb)
Supplementary Table 1. The F2 pedigrees used for QTL mapping. Parents are listed only for F1 and F2 individuals.
10519_2016_9815_MOESM2_ESM.xlsx (37 kb)
Supplementary Table 2. Contribution of each of tame and aggressive grandparent to the F2 generation.
10519_2016_9815_MOESM3_ESM.xlsx (84 kb)
Supplementary Table 3. Microsatellite markers used for genotyping fox F2 pedigrees and their locations on the meiotic linkage map (LOD 0.0).
10519_2016_9815_MOESM4_ESM.docx (145 kb)
Supplementary Table 4. List of traits used for PC analysis and frequency of trait observations in aggressive, tame, F1, and F2 populations (A) and average PC values in parental and cross-bred populations (B). For aggressive, tame, and F1 populations frequency of the trait observations in the test #1 are listed, for the F2 population, mean of frequency observations in the tests #1 and #2 is presented. The standard test included four steps: Step A - observer stands calmly near the closed cage but does not deliberately try to attract the fox’s attention; Step B - observer opens the cage door, remains nearby but does not initiate any contact with the fox; Step C - observer attempts to touch the fox; Step D - observer closes the cage door, then stays calmly near the closed cage. The zones 1 and 2 are located in the front of the cage, zones 5 and 6 are located in the back of the cage, and zone 3 and 4 are in the middle. The zone 2 and 2a are the closest to a human experimenter. Tame = tame population; Aggr = aggressive population, BCT = backcross-to-tame, BCA = backcross-to-aggressive.
10519_2016_9815_MOESM5_ESM.xlsx (53 kb)
Supplementary Table 5. Trait loadings in the first three PCs calculated for each individual test step. The traits with highest absolute loadings to the corresponding PC are highlighted by green (top 20th percentile) and blue (lowest 20th percentile).
10519_2016_9815_MOESM6_ESM.docx (66 kb)
Supplementary Table 6. Estimated additive (a) and dominance (d) effects of QTL identified for PC1 traits and estimation of population differences and residual variance explained by each QTL. Tame = tame population; Aggr = aggressive population.
10519_2016_9815_MOESM7_ESM.docx (42 kb)
Supplementary Table 7. Correlation coefficients for pairs of PC1 phenotypes in parental and experimental populations. BCT = backcross-to-tame strain; BCA = backcross-to-aggressive strain.
10519_2016_9815_MOESM8_ESM.doc (158 kb)
Supplementary Figure 1. Box plots of first three PCs calculated for each individual test step. Horizontal bars within the boxes indicate the population median. The bottom and top edges of the boxes indicate the 25 and 75 percentiles. The whiskers indicate the range of data up to 1.5 times the interquartile range. Outliers are shown as individual circles. Data for four populations Tame = tame, Aggr = aggressive, F1 and F2 are shown.
10519_2016_9815_MOESM9_ESM.doc (7.9 mb)
Supplementary Figure 2. Histogram of the first three PCs calculated for each individual test step. Data for F2 population are shown.
10519_2016_9815_MOESM10_ESM.xls (260 kb)
Supplementary Figure 3. Heat map of 98 traits scored in the standard test. Spearman Rank correlation of 98 traits was calculated for all 1287 individuals included in PC analysis. The traits are ordered using two parameters: 1) test step; 2) correlation coefficient with step specific PC1 phenotype.
10519_2016_9815_MOESM11_ESM.pdf (151 kb)
Supplementary Figures 4. Main effect QTL for PC phenotypes. QTL plot for each PC phenotype with significant (p = 0.05) and suggested (p ≈ 0.20) QTL thresholds indicated. Vertical dashed lines indicate boundaries of fox autosomes.
10519_2016_9815_MOESM12_ESM.pdf (1.2 mb)
Supplementary Figure 5. QTL plot for each trait. QTL plot for each trait with genome-wide significance threshold (p=0.05) indicated (F-value = 8.3). See also Table 4.
10519_2016_9815_MOESM13_ESM.pdf (1.5 mb)
Supplementary Figures 6. QTL plot for each PC and the associated traits. QTL plots for each of the PC phenotypes are in blue. Associated traits in the 20th percentile (see Table 2 for details) indicated in red (TL-20). Genome-wide significance threshold (p=0.05) is indicated (F-value of 8.3). See also Supplementary Figures 4 and 5 for individual PC and trait profiles.
10519_2016_9815_MOESM14_ESM.pdf (175 kb)
Supplementary Figure 7. Correlation between B.PC1 and C.PC1 phenotypes in tame, aggressive, backcross-to-tame, backcross-to-aggressive, and F2 populations.
10519_2016_9815_MOESM15_ESM.pdf (33 kb)
Supplementary Figure 8. Genotype-phenotype map for C.PC1 on VVU1. Genotype-phenotype map and associated QTL position on chromosome 1 (65 cM) for the phenotype C.PC1 and traits with significant contribution to C.PC1 (Table 2). The QTL plot indicates a position in the genome for which the association between phenotype and genotype is presented (vertical red line). The box-plot indicates the C.PC1 phenotype values for the three genotypic classes (TT, TA, and AA). The stacked bar graph for significant traits demonstrates a relative proportion of F2 individuals from the three genotypic classes (TT, TA and AA) which did not show the trait (0) demonstrated it in one of the two tests (0.5), or in both tests analyzed (1).
10519_2016_9815_MOESM16_ESM.pdf (30 kb)
Supplementary Figure 9. Genotype-phenotype map for C.PC1 on VVU5. Genotype-phenotype map and associated QTL position on chromosome 1 (65 cM) for the phenotype C.PC1 and traits with significant contribution to C.PC1 (Table 2). The QTL plot indicates a position in the genome for which the association between phenotype and genotype is presented (vertical red line). The box-plot indicates the C.PC1 phenotype values for the three genotypic classes (TT, TA, and AA). The stacked bar graph for significant traits demonstrates a relative proportion of F2 individuals from the three genotypic classes (TT, TA and AA) which did not show the trait (0) demonstrated it in one of the two tests (0.5), or in both tests analyzed (1).
10519_2016_9815_MOESM17_ESM.pdf (2.2 mb)
Supplementary Figure 10. Interacting pairs of loci for B.PC1 and C.PC1 phenotypes. Main effect QTL for B.PC1 and C.PC1 in the fox genome indicated on the X- and Y-axes with a genome-wide significance threshold of 5%. 2D plot of SSE (residual sum of squares) of all epistatic pairs (low values darker). Significant interacting pairs indicated by blue dots. See Table 5 for details.
10519_2016_9815_MOESM18_ESM.docx (149 kb)
Supplementary File 1. R code used in simulation study.


  1. Albert FW et al (2009) Genetic architecture of tameness in a rat model of animal domestication. Genetics 182(2):541–554CrossRefPubMedPubMedCentralGoogle Scholar
  2. Anholt RRH, Mackay TFC (2009) Principles of behavioral genetics, 1st edn. Academic Press, OxfordGoogle Scholar
  3. Axelsson E et al (2013) The genomic signature of dog domestication reveals adaptation to a starch-rich diet. Nature 495(7441):360–364CrossRefPubMedGoogle Scholar
  4. Barrett CE et al (2013) Variation in vasopressin receptor (Avpr1a) expression creates diversity in behaviors related to monogamy in prairie voles. Horm Behav 63(3):518–526CrossRefPubMedPubMedCentralGoogle Scholar
  5. Bell AM (2005) Behavioral differences between individuals and populations of threespined stickleback. J Evol Biol 18(2):464–473CrossRefPubMedGoogle Scholar
  6. Belyaev DK (1979) Destabilizing selection as a factor in domestication. J Hered 70(5):301–308PubMedGoogle Scholar
  7. Brent LJ et al (2013) Genetic origins of social networks in rhesus macaques. Sci Rep 3:1042CrossRefPubMedPubMedCentralGoogle Scholar
  8. Brodkin ES et al (2002) Identification of quantitative trait loci that affect aggressive behavior in mice. J Neurosci 22(3):1165–1170PubMedGoogle Scholar
  9. Carlborg Ö, Andersson L (2002) Use of randomization testing to detect multiple epistatic QTLs. Genet Res 79(2):175–184CrossRefPubMedGoogle Scholar
  10. Carlborg Ö, Andersson L, Kinghorn B (2000) The use of a genetic algorithm for simultaneous mapping of multiple interacting quantitative trait loci. Genetics 155(4):2003–2010PubMedPubMedCentralGoogle Scholar
  11. Carlborg Ö et al (2003) A global search reveals epistatic interaction between QTL for early growth in the chicken. Genome Res 13(3):413–421CrossRefPubMedPubMedCentralGoogle Scholar
  12. Carneiro M et al (2014) Rabbit genome analysis reveals a polygenic basis for phenotypic change during domestication. Science 345(6200):1074–1079CrossRefPubMedGoogle Scholar
  13. Champoux M, Higley JD, Suomi SJ (1997) Behavioral and physiological characteristics of Indian and Chinese-Indian hybrid rhesus macaque infants. Dev Psychobiol 31(1):49–63CrossRefPubMedGoogle Scholar
  14. Churchill GA, Doerge RW (1994) Empirical threshold values for quantitative trait mapping. Genetics 138(3):963–971PubMedPubMedCentralGoogle Scholar
  15. Crooks L, Nettelblad C, Carlborg Ö (2011) An improved method for estimating chromosomal line origin in QTL analysis of crosses between outbred lines. G3: Genes| Genomes| Genetics 1(1):57–64CrossRefPubMedCentralGoogle Scholar
  16. Dow HC et al (2011) Genetic dissection of intermale aggressive behavior in BALB/cJ and A/J mice. Genes Brain Behav 10(1):57–68CrossRefPubMedGoogle Scholar
  17. Driscoll CA et al (2007) The Near Eastern origin of cat domestication. Science 317(5837):519–523CrossRefPubMedGoogle Scholar
  18. Fairbanks LA et al (2004) Genetic contributions to social impulsivity and aggressiveness in vervet monkeys. Biol Psychiatry 55(6):642–647CrossRefPubMedGoogle Scholar
  19. Freudenberg F et al (2016) Aggression in non-human vertebrates: genetic mechanisms and molecular pathways. Am J Med Genet Part B Neuropsychiatr Genet 171(5):603–640CrossRefGoogle Scholar
  20. Gilbert JR, Vance JM (1994) Isolation of genomic DNA from mammalian cells. In: Dracopoli NC (ed) Current protocols in human genetics. John Wiley and Sons, New York, Appendix A.3B pp 1–6Google Scholar
  21. Green P, Fall K, Crooks S (1990) Documentation for CRI-MAP, version 2.4. Washington University School of Medicine, St. Louis, MOGoogle Scholar
  22. Groenen MA (2016) A decade of pig genome sequencing: a window on pig domestication and evolution. Genet Sel Evol 48:23CrossRefPubMedPubMedCentralGoogle Scholar
  23. Hare B et al (2005) Social cognitive evolution in captive foxes is a correlated by-product of experimental domestication. Curr Biol 15(3):226–230CrossRefPubMedGoogle Scholar
  24. Heyne HO et al (2014) Genetic influences on brain gene expression in rats selected for tameness and aggression. Genetics 198(3):1277–1290CrossRefPubMedPubMedCentralGoogle Scholar
  25. Hu Y et al (2014) Earliest evidence for commensal processes of cat domestication. Proc Natl Acad Sci USA 111(1):116–120CrossRefPubMedGoogle Scholar
  26. Johnson JL et al (2015) Genotyping-By-Sequencing (GBS) detects genetic structure and confirms behavioral QTL in tame and aggressive foxes (Vulpes vulpes). PLoS ONE 10(6):e0127013CrossRefPubMedPubMedCentralGoogle Scholar
  27. King LB et al (2016) Variation in the oxytocin receptor gene predicts brain region-specific expression and social attachment. Biol Psychiatry 80(2):160–169CrossRefPubMedGoogle Scholar
  28. Kukekova AV et al (2004) A marker set for construction of a genetic map of the silver fox (Vulpes vulpes). J Hered 95(3):185–194CrossRefPubMedGoogle Scholar
  29. Kukekova AV et al (2007) A meiotic linkage map of the silver fox, aligned and compared to the canine genome. Genome Res 17(3):387–399CrossRefPubMedPubMedCentralGoogle Scholar
  30. Kukekova AV et al (2008) Measurement of segregating behaviors in experimental silver fox pedigrees. Behav Genet 38(2):185–194CrossRefPubMedGoogle Scholar
  31. Kukekova AV et al (2011) Mapping Loci for fox domestication: deconstruction/reconstruction of a behavioral phenotype. Behav Genet 41(4):593–606CrossRefPubMedGoogle Scholar
  32. Kukekova AV et al (2012) Genetics of behavior in the silver fox. Mamm Genome 23(1–2):164–177CrossRefPubMedGoogle Scholar
  33. Kukekova AV, Trut LN, Acland GM (2014) Genetics of domesticated behavior in dogs and foxes. In: Grandin T, Deesing MJ (eds) Genetics and the Behavior of Domestic Animals, 2nd edn. Academic Press, Salt Lake City, pp 361–396CrossRefGoogle Scholar
  34. Landis JR, Koch GG (1977) The measurement of observer agreement for categorical data. Biometrics 33(1):159–174CrossRefPubMedGoogle Scholar
  35. Lindblad-Toh K et al (2005) Genome sequence, comparative analysis and haplotype structure of the domestic dog. Nature 438(7069):803–819CrossRefPubMedGoogle Scholar
  36. Matise TC, Perlin M, Chakravarti A (1994) Automated construction of genetic linkage maps using an expert system (MultiMap): a human genome linkage map. Nat Genet 6(4):384–390CrossRefPubMedGoogle Scholar
  37. McGraw LA, Young LJ (2010) The prairie vole: an emerging model organism for understanding the social brain. Trends Neurosci 33(2):103–109CrossRefPubMedGoogle Scholar
  38. Montague MJ et al (2014) Comparative analysis of the domestic cat genome reveals genetic signatures underlying feline biology and domestication. Proc Natl Acad Sci USA 111(48):17230–17235CrossRefPubMedPubMedCentralGoogle Scholar
  39. Nehrenberg DL et al (2010) Genomic mapping of social behavior traits in a F2 cross derived from mice selectively bred for high aggression. BMC Genet 11:113CrossRefPubMedPubMedCentralGoogle Scholar
  40. Nelson RM et al (2013) MAPfastR: quantitative trait loci mapping in outbred line crosses. G3: Genes| Genomes| Genetics 3(12):2147–2149CrossRefPubMedPubMedCentralGoogle Scholar
  41. Ott J (1999) Analysis of Human Genetic Linkage, 3rd edn. The Johns Hopkins University Press, Baltimore and LondonGoogle Scholar
  42. Price EO (2008) Principles and applications of domestic animal behavior. CABI Publishing, New YorkGoogle Scholar
  43. Roubertoux PL et al (2005) Attack behaviors in mice: from factorial structure to quantitative trait loci mapping. Eur J Pharmacol 526(1–3):172–185CrossRefPubMedGoogle Scholar
  44. Savolainen P et al (2002) Genetic evidence for an East Asian origin of domestic dogs. Science 298(5598):1610–1613CrossRefPubMedGoogle Scholar
  45. Svartberg K, Forkman B (2002) Personality traits in the domestic dog (Canis familiaris). Appl Anim Behav Sci 79(2):133–155CrossRefGoogle Scholar
  46. Takahashi A, Shiroishi T, Koide T (2014) Genetic mapping of escalated aggression in wild-derived mouse strain MSM/Ms: association with serotonin-related genes. Front Neurosci 8:156CrossRefPubMedPubMedCentralGoogle Scholar
  47. Takahashi A et al (2015) Mapping of Genetic Factors That Elicit Intermale Aggressive Behavior on Mouse Chromosome 15: intruder Effects and the Complex Genetic Basis. PLoS ONE 10(9):e0137764CrossRefPubMedPubMedCentralGoogle Scholar
  48. Trut LN (1980) The genetics and phenogenetics of domestic behaviour. In: Belyaev DK (ed) Problems in general genetics (Proceeding of the XIV International Congress of Genetics). Mir Publishers, Moscow, pp 123–137Google Scholar
  49. Trut LN (1999) Early canid domestication: the farm-fox experiment. Am Sci 87(2):160–169CrossRefGoogle Scholar
  50. Trut L, Oskina I, Kharlamova A (2009) Animal evolution during domestication: the domesticated fox as a model. BioEssays 31(3):349–360CrossRefPubMedPubMedCentralGoogle Scholar
  51. Trut LN, Oskina IN, Kharlamova AV (2012) Experimental studies of early canid domestication. In: Ostrander EA, Ruvinsky A (eds) Genetics of the dog, 2nd edn. CAB International, Oxford, pp 12–37CrossRefGoogle Scholar
  52. vonHoldt BM et al (2010) Genome-wide SNP and haplotype analyses reveal a rich history underlying dog domestication. Nature 464(7290):898–902CrossRefPubMedPubMedCentralGoogle Scholar
  53. Wang GD et al (2013) The genomics of selection in dogs and the parallel evolution between dogs and humans. Nat Commun 4:1860CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  • Ronald M. Nelson
    • 1
    • 2
  • Svetlana V. Temnykh
    • 3
  • Jennifer L. Johnson
    • 4
  • Anastasiya V. Kharlamova
    • 5
  • Anastasiya V. Vladimirova
    • 5
  • Rimma G. Gulevich
    • 5
  • Darya V. Shepeleva
    • 5
  • Irina N. Oskina
    • 5
  • Gregory M. Acland
    • 3
  • Lars Rönnegård
    • 1
    • 6
  • Lyudmila N. Trut
    • 5
  • Örjan Carlborg
    • 1
    • 2
  • Anna V. Kukekova
    • 4
  1. 1.Division of Computational Genetics, Department of Clinical SciencesSwedish University of Agricultural SciencesUppsalaSweden
  2. 2.Department of Medical Biochemistry and MicrobiologyUppsala UniversityUppsalaSweden
  3. 3.Baker Institute for Animal Health, College of Veterinary MedicineCornell UniversityIthacaUSA
  4. 4.Animal Sciences Department, College of Agricultural, Consumer and Environmental SciencesUniversity of Illinois at Urbana-ChampaignUrbanaUSA
  5. 5.Institute of Cytology and Genetics of the Russian Academy of SciencesNovosibirskRussia
  6. 6.Section of Statistics, School of Technology and Business StudiesDalarna UniversityFalunSweden

Personalised recommendations