Insular cortex mediates approach and avoidance responses to social affective stimuli

Abstract

Social animals detect the affective states of conspecifics and utilize this information to orchestrate social interactions. In a social affective preference text in which experimental adult male rats could interact with either naive or stressed conspecifics, the experimental rats either approached or avoided the stressed conspecific, depending upon the age of the conspecific. Specifically, experimental rats approached stressed juveniles but avoided stressed adults. Inhibition of insular cortex, which is implicated in social cognition, and blockade of insular oxytocin receptors disrupted the social affective behaviors. Oxytocin application increased intrinsic excitability and synaptic efficacy in acute insular cortex slices, and insular oxytocin administration recapitulated the behaviors observed toward stressed conspecifics. Network analysis of c-Fos immunoreactivity in 29 regions identified functional connectivity between insular cortex, prefrontal cortex, amygdala and the social decision-making network. These results implicate insular cortex as a key component in the circuit underlying age-dependent social responses to stressed conspecifics.

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Fig. 1: Social affective preference (SAP).
Fig. 2: Optogenetic silencing of insular cortex during SAP tests.
Fig. 3: Intrinsic membrane properties in insular cortex pyramidal neurons are modulated by oxytocin and depend upon PKC.
Fig. 4: Oxytocin modulates excitatory synaptic transmission in the insular cortex.
Fig. 5: Social affective behaviors require insular cortex oxytocin.
Fig. 6: Insular cortex and the SDMN.

References

  1. 1.

    Darwin, C. The Expression of the Emotions in Man and Animals. (St. Martin’s Press, London, New York, 1976). Julian Friedmann.

    Google Scholar 

  2. 2.

    O’Connell, L. A. & Hofmann, H. A. The vertebrate mesolimbic reward system and social behavior network: a comparative synthesis. J. Comp. Neurol. 519, 3599–3639 (2011).

    Article  PubMed  Google Scholar 

  3. 3.

    Gu, X. et al. Anterior insular cortex is necessary for empathetic pain perception. Brain 135, 2726–2735 (2012).

    Article  PubMed  PubMed Central  Google Scholar 

  4. 4.

    Ibañez, A., Gleichgerrcht, E. & Manes, F. Clinical effects of insular damage in humans. Brain Struct. Funct. 214, 397–410 (2010).

    Article  PubMed  Google Scholar 

  5. 5.

    Leigh, R. et al. Acute lesions that impair affective empathy. Brain 136, 2539–2549 (2013).

    Article  PubMed  PubMed Central  Google Scholar 

  6. 6.

    Blanken, L. M. et al. Cortical morphology in 6- to 10-year old children with autistic traits: a population-based neuroimaging study. Am. J. Psychiatry 172, 479–486 (2015).

    Article  PubMed  Google Scholar 

  7. 7.

    Di Martino, A. et al. Relationship between cingulo-insular functional connectivity and autistic traits in neurotypical adults. Am. J. Psychiatry 166, 891–899 (2009).

    Article  PubMed  PubMed Central  Google Scholar 

  8. 8.

    Morita, T. et al. Emotional responses associated with self-face processing in individuals with autism spectrum disorders: an fMRI study. Soc. Neurosci. 7, 223–239 (2012).

    Article  PubMed  Google Scholar 

  9. 9.

    Odriozola, P. et al. Insula response and connectivity during social and non-social attention in children with autism. Soc. Cogn. Affect. Neurosci. 11, 433–444 (2016).

    Article  PubMed  Google Scholar 

  10. 10.

    Gogolla, N. The insular cortex. Curr. Biol. 27, R580–R586 (2017).

    CAS  Article  PubMed  Google Scholar 

  11. 11.

    Allen, G. V. & Cechetto, D. F. Functional and anatomical organization of cardiovascular pressor and depressor sites in the lateral hypothalamic area. II. Ascending projections. J. Comp. Neurol. 330, 421–438 (1993).

  12. 12.

    Ohara, P. T. et al. Dopaminergic input to GABAergic neurons in the rostral agranular insular cortex of the rat. J. Neurocytol. 32, 131–141 (2003).

    CAS  Article  PubMed  Google Scholar 

  13. 13.

    Shi, C. J. & Cassell, M. D. Cortical, thalamic, and amygdaloid connections of the anterior and posterior insular cortices. J. Comp. Neurol. 399, 440–468 (1998).

    CAS  Article  PubMed  Google Scholar 

  14. 14.

    Sato, F. et al. Projections from the insular cortex to pain-receptive trigeminal caudal subnucleus (medullary dorsal horn) and other lower brainstem areas in rats. Neuroscience 233, 9–27 (2013).

    CAS  Article  PubMed  Google Scholar 

  15. 15.

    Allen, G. V., Saper, C. B., Hurley, K. M. & Cechetto, D. F. Organization of visceral and limbic connections in the insular cortex of the rat. J. Comp. Neurol. 311, 1–16 (1991).

    CAS  Article  PubMed  Google Scholar 

  16. 16.

    Yasui, Y., Breder, C. D., Saper, C. B. & Cechetto, D. F. Autonomic responses and efferent pathways from the insular cortex in the rat. J. Comp. Neurol. 303, 355–374 (1991).

    CAS  Article  PubMed  Google Scholar 

  17. 17.

    Knobloch, H. S. et al. Evoked axonal oxytocin release in the central amygdala attenuates fear response. Neuron 73, 553–566 (2012).

    CAS  Article  PubMed  Google Scholar 

  18. 18.

    Dumais, K. M., Bredewold, R., Mayer, T. E. & Veenema, A. H. Sex differences in oxytocin receptor binding in forebrain regions: correlations with social interest in brain region- and sex- specific ways. Horm. Behav. 64, 693–701 (2013).

    CAS  Article  PubMed  Google Scholar 

  19. 19.

    Donaldson, Z. R. & Young, L. J. Oxytocin, vasopressin, and the neurogenetics of sociality. Science 322, 900–904 (2008).

    CAS  Article  PubMed  Google Scholar 

  20. 20.

    Shamay-Tsoory, S. G. & Abu-Akel, A. The social salience hypothesis of oxytocin. Biol. Psychiatry 79, 194–202 (2016).

    CAS  Article  PubMed  Google Scholar 

  21. 21.

    Burkett, J. P. et al. Oxytocin-dependent consolation behavior in rodents. Science 351, 375–378 (2016).

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  22. 22.

    Aoki, Y. et al. Oxytocin improves behavioural and neural deficits in inferring others’ social emotions in autism. Brain 137, 3073–3086 (2014).

    Article  PubMed  Google Scholar 

  23. 23.

    Scheele, D. et al. An oxytocin-induced facilitation of neural and emotional responses to social touch correlates inversely with autism traits. Neuropsychopharmacology 39, 2078–2085 (2014).

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  24. 24.

    de Waal, F. B. M. & Preston, S. D. Mammalian empathy: behavioural manifestations and neural basis. Nat. Rev. Neurosci. 18, 498–509 (2017).

    Article  PubMed  Google Scholar 

  25. 25.

    Silberberg, A. et al. Desire for social contact, not empathy, may explain “rescue” behavior in rats. Anim. Cogn. 17, 609–618 (2014).

    Article  PubMed  Google Scholar 

  26. 26.

    Bullmore, E. & Sporns, O. Complex brain networks: graph theoretical analysis of structural and functional systems. Nat. Rev. Neurosci. 10, 186–198 (2009).

    CAS  Article  PubMed  Google Scholar 

  27. 27.

    Staub, E. Helping a distressed person: social, personality, and stimulus determinants. Adv. Exp. Soc. Psychol.. 7, 293–341 (1974).

    Article  Google Scholar 

  28. 28.

    Spear, L. P. Adolescent alcohol exposure: are there separable vulnerable periods within adolescence? Physiol. Behav. 148, 122–130 (2015).

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  29. 29.

    Brudzynski, S. M. Ethotransmission: communication of emotional states through ultrasonic vocalization in rats. Curr. Opin. Neurobiol. 23, 310–317 (2013).

    CAS  Article  PubMed  Google Scholar 

  30. 30.

    Marlin, B. J., Mitre, M., D’amour, J. A., Chao, M. V. & Froemke, R. C. Oxytocin enables maternal behaviour by balancing cortical inhibition. Nature 520, 499–504 (2015).

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  31. 31.

    Gimpl, G. & Fahrenholz, F. The oxytocin receptor system: structure, function, and regulation. Physiol. Rev. 81, 629–683 (2001).

    CAS  Article  PubMed  Google Scholar 

  32. 32.

    Power, J. D., Schlaggar, B. L., Lessov-Schlaggar, C. N. & Petersen, S. E. Evidence for hubs in human functional brain networks. Neuron 79, 798–813 (2013).

    CAS  Article  PubMed  Google Scholar 

  33. 33.

    Mikosz, M., Nowak, A., Werka, T. & Knapska, E. Sex differences in social modulation of learning in rats. Sci. Rep. 5, 18114 (2015).

  34. 34.

    Choe, H. K. et al. Oxytocin mediates entrainment of sensory stimuli to social cues of opposing valence. Neuron 87, 152–163 (2015).

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  35. 35.

    Insel, T. R. & Fernald, R. D. How the brain processes social information: searching for the social brain. Annu. Rev. Neurosci. 27, 697–722 (2004).

    CAS  Article  PubMed  Google Scholar 

  36. 36.

    Valenta, J. G. & Rigby, M. K. Discrimination of the odor of stressed rats. Science 161, 599–601 (1968).

    CAS  Article  PubMed  Google Scholar 

  37. 37.

    Sotocinal, S. G. et al. The Rat Grimace Scale: a partially automated method for quantifying pain in the laboratory rat via facial expressions. Mol. Pain. 7, 55 (2011).

    PubMed  PubMed Central  Google Scholar 

  38. 38.

    Kiyokawa, Y. Social odors: alarm pheromones and social buffering. Curr. Top. Behav. Neurosci. 30, 47–65 (2017).

    Article  PubMed  Google Scholar 

  39. 39.

    Burgdorf, J. et al. Ultrasonic vocalizations of rats (Rattus norvegicus) during mating, play, and aggression: behavioral concomitants, relationship to reward, and self-administration of playback. J. Comp. Psychol. 122, 357–367 (2008).

    Article  PubMed  Google Scholar 

  40. 40.

    Oettl, L.L. & Kelsch, W. Oxytocin and olfaction. Curr. Top. Behav. Neurosci. (2017).

  41. 41.

    Langford, D. J. et al. Social approach to pain in laboratory mice. Soc. Neurosci. 5, 163–170 (2010).

    Article  PubMed  Google Scholar 

  42. 42.

    Ben-Ami Bartal, I., Rodgers, D. A., Bernardez Sarria, M. S., Decety, J. & Mason, P. Pro-social behavior in rats is modulated by social experience. eLife 3, e01385 (2014).

    Article  PubMed  PubMed Central  Google Scholar 

  43. 43.

    de Waal, F. B. Putting the altruism back into altruism: the evolution of empathy. Annu. Rev. Psychol. 59, 279–300 (2008).

    Article  PubMed  Google Scholar 

  44. 44.

    Moaddab, M., Hyland, B. I. & Brown, C. H. Oxytocin excites nucleus accumbens shell neurons in vivo. Mol. Cell. Neurosci. 68, 323–330 (2015).

    CAS  Article  PubMed  Google Scholar 

  45. 45.

    Adolphs, R. The social brain: neural basis of social knowledge. Annu. Rev. Psychol. 60, 693–716 (2009).

    Article  PubMed  PubMed Central  Google Scholar 

  46. 46.

    Bartz, J. A., Zaki, J., Bolger, N. & Ochsner, K. N. Social effects of oxytocin in humans: context and person matter. Trends Cogn. Sci. 15, 301–309 (2011).

    CAS  PubMed  Google Scholar 

  47. 47.

    Decety, J. & Moriguchi, Y. The empathic brain and its dysfunction in psychiatric populations: implications for intervention across different clinical conditions. Biopsychosoc. Med.. 1, 22 (2007).

    Article  PubMed  PubMed Central  Google Scholar 

  48. 48.

    Thioux, M. & Keysers, C. Empathy: shared circuits and their dysfunctions. Dialog-. Clin. Neurosci. 12, 546–552 (2010).

    Google Scholar 

  49. 49.

    LoParo, D. & Waldman, I. D. The oxytocin receptor gene (OXTR) is associated with autism spectrum disorder: a meta-analysis. Mol. Psychiatry 20, 640–646 (2015).

    CAS  Article  PubMed  Google Scholar 

  50. 50.

    Skuse, D. H. et al. Common polymorphism in the oxytocin receptor gene (OXTR) is associated with human social recognition skills. Proc. Natl. Acad. Sci. USA 111, 1987–1992 (2014).

    CAS  Article  PubMed  Google Scholar 

  51. 51.

    Paxinos, G. & Watson, C. The Rat Brain Atlas in Stereotaxic Coordinates. 4th edition, (Academic Press, San Diego, CA, USA, 1998).

    Google Scholar 

  52. 52.

    Smith, C. J., Wilkins, K. B., Mogavero, J. N. & Veenema, A. H. Social novelty investigation in the juvenile rat: modulation by the μ-opioid system. J. Neuroendocrinol. 27, 752–764 (2015).

    CAS  Article  PubMed  Google Scholar 

  53. 53.

    Christianson, J. P. et al. Safety signals mitigate the consequences of uncontrollable stress via a circuit involving the sensory insular cortex and bed nucleus of the stria terminalis. Biol. Psychiatry 70, 458–464 (2011).

    Article  PubMed  PubMed Central  Google Scholar 

  54. 54.

    Manning, M. et al. Oxytocin and vasopressin agonists and antagonists as research tools and potential therapeutics. J. Neuroendocrinol. 24, 609–628 (2012).

  55. 55.

    Lukas, M., Toth, I., Veenema, A. H. & Neumann, I. D. Oxytocin mediates rodent social memory within the lateral septum and the medial amygdala depending on the relevance of the social stimulus: male juvenile versus female adult conspecifics. Psychoneuroendocrinology 38, 916–926 (2013).

    CAS  Article  PubMed  Google Scholar 

  56. 56.

    Tye, K. M. et al. Amygdala circuitry mediating reversible and bidirectional control of anxiety. Nature 471, 358–362 (2011).

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  57. 57.

    Sidiropoulou, K. et al. Dopamine modulates an mGluR5-mediated depolarization underlying prefrontal persistent activity. Nat. Neurosci. 12, 190–199 (2009).

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  58. 58.

    Dölen, G., Darvishzadeh, A., Huang, K. W. & Malenka, R. C. Social reward requires coordinated activity of nucleus accumbens oxytocin and serotonin. Nature 501, 179–184 (2013).

    Article  PubMed  PubMed Central  Google Scholar 

  59. 59.

    Szot, P., Bale, T. L. & Dorsa, D. M. Distribution of messenger RNA for the vasopressin V1a receptor in the CNS of male and female rats. Brain Res. Mol. Brain Res. 24, 1–10 (1994).

    CAS  Article  PubMed  Google Scholar 

  60. 60.

    Dumais, K. M. & Veenema, A. H. Vasopressin and oxytocin receptor systems in the brain: Sex differences and sex-specific regulation of social behavior. Front. Neuroendocrinol. 40, 1–23 (2016).

    CAS  Article  PubMed  Google Scholar 

  61. 61.

    Varela, J. A., Wang, J., Christianson, J. P., Maier, S. F. & Cooper, D. C. Control over stress, but not stress per se increases prefrontal cortical pyramidal neuron excitability. J. Neurosci. 32, 12848–12853 (2012).

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  62. 62.

    Turrigiano, G. G., Leslie, K. R., Desai, N. S., Rutherford, L. C. & Nelson, S. B. Activity-dependent scaling of quantal amplitude in neocortical neurons. Nature 391, 892–896 (1998).

    CAS  Article  PubMed  Google Scholar 

  63. 63.

    Gschwendt, M. et al. Inhibition of protein kinase C mu by various inhibitors. Differentiation from protein kinase c isoenzymes. FEBS Lett.. 392, 77–80 (1996).

    CAS  Article  PubMed  Google Scholar 

  64. 64.

    Christianson, J. P. et al. The sensory insular cortex mediates the stress-buffering effects of safety signals but not behavioral control. J. Neurosci. 28, 13703–13711 (2008).

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  65. 65.

    Vetere, G. et al. Chemogenetic interrogation of a brain-wide fear memory network in mice. Neuron 94, 363–374.e4 (2017).

    CAS  Article  PubMed  Google Scholar 

  66. 66.

    Wheeler, A. L. et al. Identification of a functional connectome for long-term fear memory in mice. PLOS Comput. Biol. 9, e1002853 (2013).

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  67. 67.

    Tanimizu, T. et al. Functional connectivity of multiple brain regions required for the consolidation of social recognition memory. J. Neurosci. 37, 4103–4116 (2017).

    CAS  Article  PubMed  Google Scholar 

  68. 68.

    Teles, M. C., Almeida, O., Lopes, J. S. & Oliveira, R. F. Social interactions elicit rapid shifts in functional connectivity in the social decision-making network of zebrafish. Proc. Biol. Sci.. 282, 20151099 (2015).

    Article  PubMed  PubMed Central  Google Scholar 

  69. 69.

    Newman, M. E. Modularity and community structure in networks. Proc. Natl. Acad. Sci. USA 103, 8577–8582 (2006).

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  70. 70.

    Wickham, H. ggplot2: Elegant Graphics for Data Analysis. (Springer-Verlag, New York, 2009).

    Google Scholar 

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Acknowledgements

The authors thank M. Manning (University of Toledo) for providing OTRa, K. Deisseroth (Stanford University) for making optogenetic vectors freely available, and A. Veenema and D. Adams for discussions related to the project. Funding for this work was provided by the Boston College Undergraduate Research Fellowships; National Science Foundation Grant #1258923 to M.M.R.-C; NIH grants MH103401 to M.R. and MH093412 and MH109545 to J.P.C.; and Brain and Behavior Research Foundation grant No. 19417 to J.P.C.

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Contributions

Conceptualization: J.P.C., J.A.V., M.M.R.-C., M.R. Methodology: M.M.R.-C., J.A.V., M.R. J.P.C. Investigation: J.A.V., M.M.R.-C., K.B.G., A.P., M.M., M.R., J.P.C. Writing, original draft: J.A.V., M.M.R.-C., and J.P.C.;;revising & editing: M.M.R.-C., M.R., J.P.C. Funding acquisition, J.P.C., M.M.R.-C.

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Correspondence to John P. Christianson.

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Integrated supplementary information

Supplementary Figure 1 Conspecific behavior in social affective preference test.

(a) An observer blind to conspecific treatment tallied the time spent inactive, self-grooming, engaged in social interaction with the experimental adult and time spent sniffing and exploring the conspecific chamber of 12 pairs of target rats from each of the PN 30 and PN 50 treatment groups. The average portion (% of total) time spent in each behavior during the 5-minute test is shown for each treatment. The analyses indicate that the conspecifics spent the majority of time in active behaviors, namely sniffing and exploring the chamber. (b) Mean (+/− SEM) time spent self-grooming. Separate t-tests were conducted to compare time in each behavior and revealed a significant increase in self grooming after stress in the PN 30 group. *t(22) = 2.47, p = 0.022 (2-tailed). No other comparisons reached significance, and self grooming did not correlate with behavior of the experimental adult rats.

Supplementary Figure 2 Repeated one-on-one testing and SAP tests in females.

(a) Adult male rats (n = 6) were given a series of 4, 1-on-1 social interaction tests (5 min duration) in each of the following conditions: naive-juvenile, naive-adult, stress-juvenile and stress-adult; test order was counter balanced using a Latin square design. Tests were exactly as described in the Online Methods and separated by 20 to 30 min. All conspecific stimuli were novel/unfamiliar pairs. Two way repeated measures ANOVA revealed significant interaction of age and stress, F(1, 5) = 20.549, p = 0.006, with significant differences between Naive and Stress at both ages, *p = 0.022 (2-tailed), #p = 0.028 (1-tailed). (b) Mean (+/− SEM, n = 6) percent preference for stressed conspecific. This series of tests in (a) was repeated for 3 consecutive days and data were converted to percent preference for the stressed conspecific by age, and by day. PN30 and PN50 preference scores were significantly different on all three days. ANOVA revealed significant Age & Day interaction F(2, 10) = 5.211, p = 0.028, ****p < 0.0001, **p < 0.007 (Sidak). (c) Mean social interaction time. SAP tests were performed on adult female, regularly cycling rats with pairs of male juvenile or adult conspecifics as interaction stimuli (N = 8 females). SAP tests were exactly as described in the online methods. A significant 2-way interaction, F(1, 14) = 15.52, p = 0.002 indicated that the females also exhibit preference to interact with stressed juvenile conspecifics but avoid stressed adults, *p < 0.05 (Sidak). (d) Results in c. shown as Mean % time interacting with stressed conspecific for comparison, **t(7) = 3.51, p = 0.009 (2-tailed).

Supplementary Figure 3 Ultrasonic vocalizations and social interaction.

Extended analysis of data described in Fig. 1i–k. Peri-event histograms showing frequency of Rising (A) and Trill (B) USVs in the different conspecific treatment conditions (NJ, n = 10, NA, n = 10, SJ, n = 11, SA, n = 11). Time 0 is equal to the beginning of a social interaction bout. Bins are 250 ms. Calls were included from all rats in the treatment condition for social interaction bouts 3 s or longer. (C) Mean (+/− S.E.M.) rate of vocalization (rising and trills combined) 2.5 s before and 2.5 s after the beginning of a 3 s (or longer) social interaction bout. Values normalized to a pre-bout baseline 5 s prior to the social interaction. (D) Mean (+ S.E.M.) percent of total USVs detected during 5 minute social interaction bout that occurred during social interactions. Stress reduced within-bout vocalizations in the adult condition (Main effect of Age, F(3, 74) = 8.523, p < 0.0001). *SA vs. NJ, p = 0.019; SA vs. NA, p = 0.016; SA vs. SJ, p = 0.0005 (Fisher LSD). NJ = naive juvenile, NA = naive adult, SJ = stressed juvenile, SA = stressed adult.

Supplementary Figure 4 Inhibition of insular cortex with muscimol reverses SAP test behavior.

In a pilot study, rats were implanted with insular cortex cannula for microinjection exactly as described in the online methods. After 7 days of recovery rats received SAP tests with either PN 30 (n = 9) or PN 50 (n = 10) conspecifics. SAP tests were as in Fig. 2: Habituation to arena on Day 1, Habituation with exposure to naïve conspecifics on Day 2, and SAP tests with one conspecific exposed to 2 footshocks prior to test on Day 3. Either Vehicle (0.9% saline) or muscimol (100 ng/side in saline) was injected to the insular cortex 60 minutes prior to SAP tests on day 3. This experiment was different in design from the others included in the main text because rats received only 1 SAP test precluding the more powerful within-subject comparisons of drug effect. SAP behavior is shown as the mean (+/− SEM) percent of time spent interacting with the stressed conspecific of the total time spent in social interaction during the SAP test. A 2-way between subjects ANOVA revealed a significant Age by Drug interaction, F(1, 34) = 16.27, p < 0.001. Post hoc comparisons between drug conditions within each age revealed a significant decrease in time spent interacting with the stressed PN 30 juvenile conspecific by muscimol (**p = 0.005, Sidak) and a significant increase in time spent interacting with the stressed PN 50 adult conspecific (*p = 0.043, Sidak).

Supplementary Figure 5 Verification of halorhodopsin transductions and behavioral analysis in SAP tests.

Placement of AAV5-CamKIIa-eNpHr3.0-mCherry transfections in experimental adult rats given SAP tests with (a) PN 30 juvenile conspecifics or (b) PN 50 conspecifics. Different colored traces correspond to individual replicates. In each case the tip of the fiber optic cannula was found within the insular cortex and either directly above or within the area marked as containing mCherry (unamplified) fluorescence. (c) Viral transfer was validated in whole cell recordings of mCherry (above DIC image, below mCherry false colored yellow. 40x, Scale bar = 40 μm) positive neurons in acute slices containing the insular cortex. Application of green light (wavelength = 532 nm, 10 mW/mm2) through the objective of the electrophysiology microscope induced robust hyperpolarizations which silenced spiking when provided during a train of evoked spikes, scale bar 50 mV/250 ms. (d) Mean (+/− SEM) spikes during sequence of OFF/ON/OFF application as in panel (D), n = 6 cells, one-way repeated measures ANOVA, F(2, 15) = 22.82, p < 0.0001 with the spike rate in the ON condition significantly less than either off condition, ***p < 0.0001 (Sidak). (e) Mean (+SEM) time spent in different behaviors of adult conspecifics in optogenetic experiment (PN30, n = 9, PN50, n = 11). Behavior levels are equal between light ON and OFF conditions except for increased immobility in the PN50 light ON condition, Age by Light by Behavior interaction, F(4, 60) = 3.01, p = 0.025. *p = 0.001 (Sidak). The increase in immobility is likely due to 2 of 11 rats that exhibited very high immobility (25–26 s). Brain Atlas illustrations were reproduced with permission as previously published in The Brain Atlas in Stereotaxic Coordinates, 4th Edition, Paxinos, G. & Watson, C. Pages 296, 303, 306–317 & 332. Copyright Elsevier (1998).

Supplementary Figure 6 Placement of cannula tips for microinjection experiments and excluded subject data.

(a) Cannula tip placement within the insular cortex was verified in 40 μm fresh frozen sections stained with Cresyl Violet under light microscopy. Tip locations are indicated. Section distance from Bregma indicated in mm. Open/White Circles relate to Fig. 5b-c, Red circles relate to Fig. 5e and Gold circles to Fig. 5f, Grey circles related to Fig. 5g-h. (b) SAP test behavior from the rats excluded from mechanistic experiments because the baseline behavior (light OFF or vehicle condition) did not reflect the phenomena under study. LEFT: excluded from Fig. 2f-g, CENTER: excluded from Fig. 5b-c, RIGHT: excluded from Fig. 5g-h. Brain Atlas illustrations were reproduced with permission as previously published in The Brain Atlas in Stereotaxic Coordinates, 4th Edition, Paxinos, G. & Watson, C. Pages 296, 303, 306–317 & 332. Copyright Elsevier (1998).

Supplementary Figure 7 Representative examples of c-Fos immunohistochemistry and regions of interest.

Coronal diagrams on left correspond to regions of interest indicated within the representative digital photomicrographs on the right. Images were acquired and analyzed with equal exposure and analysis was conducted on images without adjustments to brightness or contrast. The representative images have been adjusted to facilitate presentation. This experiment was replicated 11 times per group. List of abbreviations is provided in Supplementary Figure 8. Scale bars = 200 μm. Brain Atlas illustrations were reproduced with permission as previously published in The Brain Atlas in Stereotaxic Coordinates, 4th Edition, Paxinos, G. & Watson, C. Pages 296, 303, 306–317 & 332. Copyright Elsevier (1998).

Supplementary Figure 8 Extended analysis of Fos network.

(A) Correlation matrices (Kendall’s tau) organized as in Fig. 6 by conspecific age and stress condition (n = 11/group, N = 44 for All condition). (B) Correlations (Box plots range = 25th to 75th percentile, line indicates median) within and between modules identified by a community detection analysis (described in Fig. 6b). (C) Effect of stress on participation coefficient computed as in Fig. 6 (ALL condition) and for each age and stress condition. NJ = naive juvenile, NA = naive adult, SJ = stressed juvenile, SA = stressed adult. See Sup. Table 1 for abbreviation key.

Supplementary Figure 9 Manipulations of insular cortex reverse SAP test behavior.

From visual inspection of the results of the optogenetic and pharmacology experiments it appeared that manipulating the insular cortex not only prevented the expression of preference behavior in the SAP test, but that it might actually switch preference in the opposite direction. To test this possibility, % time interacting with the stressed conspecific scores were pooled from all of the mechanistic experiments (halorhodopsin, OTRa, PKC inhibitor, n = 26 for PN30, n = 25 for PN50) and we conducted one-sample t-tests (2-tailed) comparing each condition to 50% (no preference). Indeed, disrupting insular function reversed SAP behavior. In PN30 rats, the mean (+SEM) % time interacting with the stressed conspecific switched away from the stressed juvenile, t(25) = 3.35, **p = 0.003 and in PN 50 rats it switched toward interacting with the stressed adult, t(24) = 2.30, *p = 0.013.

Supplementary Figure 10 Intra-insula oxytocin receptor antagonist (OTRa) does not alter social interaction.

Adult male rats were implanted with insular cortex cannula and given 3 minute social interaction tests with naive, juvenile conspecifics (N = 16). Vehicle or OTRa (as described in Online Methods) was injected 15 min before testing on day 1 or day 2. All rats were tested under both drug conditions with test order counterbalanced. There was no effect of OTRa on social interaction t(15) = 1.61, p = 0.13 (2-tailed).

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Supplementary Video 1 - Social Affective Preference Test.

In this example, the experimental adult was presented with a stressed PN 30 conspecific in the left chamber and a naïve PN 30 conspecific on the right. The appearance of a green rectangle identifies bouts of social interaction with the stressed conspecific. Video length = 39 seconds

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Rogers-Carter, M.M., Varela, J.A., Gribbons, K.B. et al. Insular cortex mediates approach and avoidance responses to social affective stimuli. Nat Neurosci 21, 404–414 (2018). https://doi.org/10.1038/s41593-018-0071-y

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