1. (Robinson, Fernald & Clayton Science, 2008) Complex Relationships Connect Genes, the Brain and Social Behavior

2. Nature and Nurture both act on the genome HeredityEnvironment Robinson Science, 2004

3. Genomics and the First Phenotype HeredityEnvironment Robinson Science, 2004

4. cGMP line graph (Ben-Shahar et al. Science, 2002) Relationship? Causal Relationship? Causal

5. Relationship? Extensive, Coherent (Whitfield et al. Science 2003) (Grozinger et al. PNAS 2003)

6. What is the Relationship Between Genes and Social Behavior? 1. 1.How responsive? 2. 2.How conservative? 3. 3.How inventive? 4. 4.New genes or new gene regulation? 5. New neurobiological insights (function, location) 6. What is the right unit of measure? Gene? Gene list? Gene regulatory network?

7. More known about bee behavior than for any other organism besides humans Normal behavior easily observed Social environment precisely controlled Recently sequenced genome Not just behaviors, but behavioral roles Why Bees?

8. Honey Bee BehavioralMaturation Nurse Forager Defenders Hive Bees

9. Flexible SociallyRegulated Research tool: Single-cohort colony

10. Hereditary and Environmental Influences on the Genome: So What? Implications for the public Implications for social science Biological implications: Pacemaker genes Pacemaker genes Ghostbuster model Ghostbuster model Potential link between behavioral evolution and gene regulation Potential link between behavioral evolution and gene regulation

11. Behavioral Maturation (Nature & Nurture) AHB/EHBAHB/EHB L/ML/M Hi/LoHi/Lo Alaux, Toth, Hunt, De Grandi-Hoffmann, Page, Amdam, LeConte cerana, florea, dorsata Sarma et al. 2007 BMC Genomicscerana, florea, dorsata Sarma et al. 2007 BMC Genomics Social regulation: precocious, normal, delayed maturation (Nurture)Social regulation: precocious, normal, delayed maturation (Nurture) (Alaux et al. 09 GBB) Circadian rhythm Bloch, Nguyen, RodriguezCircadian rhythm Bloch, Nguyen, Rodriguez Drone/worker Zayed, NguyenDrone/worker Zayed, Nguyen Q/W Nguyen, Corona, KenyonQ/W Nguyen, Corona, Kenyon Maturational pheromones (Nurture) BPBP QMP Alaux et al. 09 GBBQMP Alaux et al. 09 GBB

12. Foraging/flight-related experience dependent effects (Nurture) MB LutzMB Lutz Appointment schedule NaegerAppointment schedule Naeger Distance Sarma, Nguyen (brain regions)Distance Sarma, Nguyen (brain regions) Orientation Brockmann (central brain)Orientation Brockmann (central brain) Nutrition (Nature & Nurture) Nurse/forager (fat body)(brain from Alaux et al.)Nurse/forager (fat body)(brain from Alaux et al.) Rich/poor diet +/- QMP (brain and fat body)Rich/poor diet +/- QMP (brain and fat body) Vg RNAi (brain and fat body) Ament, Wheeler, Nguyen, Corona?Vg RNAi (brain and fat body) Ament, Wheeler, Nguyen, Corona?

13. Aggression (Nature & Nurture) AHB/EHB (guards, soldiers, foragers) alarm pheromone old/young (=nurse/forager above) Alaux, Hunt, Hoffmann, Sinha Alaux et al. PNAS Inter-individual differences in behavior (Nature & Nurture) Vibration Signalers Alaux et al. PLoS One (Schneider) Scouts Liang, Nguyen Dancers brain regions, species, old array Sarma et al. PLoS One

14. BeeSpace Experiments: Overview Foraging and Aggression: Amenable to transcriptomic analysis 20+ experiments; 2000+ microarrays; 1288 individuals Behavioral maturation (Nature & Nurture) Foraging/flight-related experience dependent effects (Nurture) Nutrition (Nature & Nurture) Aggression (Nature & Nurture) Inter-individual differences in behavior (Nature & Nurture) Gos Miklem, Jay Evans, Mark Band, Trang Nguyen, Tom Newman, Bruce Schatz, Sandra Rodriguez, Saurabh Sinha, Nathan Price Dissectors: Peir Johnson, Sophia Liang, Hadley Myers, Nick Naeger, Tom Newman, Scott Nixon, Linda Qi, Bijal Shah, Ying Wang Spotfinders: Alissa Eisenstein

15. Current Forms of Analysis of Gene Expression Results Candidate vs. PatternCandidate vs. Pattern Gene listsGene lists Localization (S. Fahrbach)Localization (S. Fahrbach) Gene list comparisonsGene list comparisons Gene clusteringGene clustering PCAPCA cis-regulatorycis-regulatory Co-expression matricesCo-expression matrices Class predictionClass prediction GO and GeneList AnnotatorGO and GeneList Annotator Genes showing species differences in regional expression (current study) 74 Behavior (GO:0007610) 23 946 73 15 241 50 Signal transduction (GO:0007165)

16. Components of Aggression: Hereditary Hereditary Environmental Environmental Maturational Maturational Behavioral Behavioral

18. P<0.001 a ac bc b Colony rearedCage reared *** (Alaux & Robinson J. Chem Ecol. 2007) Phermone exposure 0123456 Number of fanning bees 0 5 10 15 20 25 Colony-reared Cage-reared Lab (Alaux & Robinson J. Chem Ecol. 2007) Effect of Exposure to Alarm Pheromone on Bee Behavior and c-Jun Antennal Lobe Expression N = 8 colonies N = 10 bees

19. (Alaux et al. Submitted) Genome Sensitive to Social Influence: 25% Variation in Brain Gene Expression Due to Colony Genotype

20. (Alaux et al. Submitted) Nature and Nurture Converge: Gene Regulation Across Three Time Scales

21. (Alaux et al. Submitted) Nature and Nurture Converge: Decreased Brain Metabolism Associated with Aggression

22. 1. 1.New Nature-Nurture analysis provides link between gene regulation and behavioral evolution 2. 2.Killer bee molecular profile: like they’re “on” alarm pheromone, even when they are not 3. 3.Down-regulation of brain metabolism associated with behavioral arousal Conclusions from Aggression Studies

23. General Conclusions So Far Genome sensitive to social influence Social evolution via changes in gene regulation Down-regulation of brain metabolism associated with behavioral arousal Other outcomes: candidate genes for many social roles; more to come

24. Challenges How to prove causality when each behavior is associated with hundreds to thousands of changes in brain gene expression? What’s the right level of biological organization for cross- species analysis to provide meaningful insights into brain function? gene? gene regulatory network?

25. Current and Former Lab Members Cedric Alaux Seth Ament Axel Brockmann Tom Newman Trang Nguyen Karen Pruiett photo by Adam Makarenko Gloria Hoffmann (ARS Tucson) Susan Fahrbach (Wake Forest) Greg Hunt (Purdue) Cynthia Kenyon (UCSF) Yves LeConte (INRA, France) Rob Page and Gro Amdam (ASU) Sandra Rodriguez Bruce Schatz Stan Schneider (UNC-Charlotte) Saurabh Sinha Amro Zayed Collaborators Keck Center for Functional and Comparative Genomics Mark Band Mark Band Miguel Corona Christina Grozinger Sophia Liang Claudia Lutz Nick Naeger Moushumi Sarma Amy Toth Marsha Wheeler Charles Whitfield