1. Stress-induced endocrine responses and anxiety: the effects of comfort food Regina Célia Spadari 2 nd International Conference on Endocrinology Chicago - October 19-22

2. Obesity Prevalence of overweight and obesity. In the USA approximately two thirds of the population is overweight, and nearly one third is obese. The causes of the epidemic obesity are complex, and stress has been identified as an important factor. Flegal et al., 2010 Increased rates of obesity have been accompanied by a concomitant rise in perceived stress in North America.

3. Peripheral Stress System SNS NE Adrenal medulla epinephrine norepinephrine Hypothalamus (PVN) CRH Pituitary (anterior) ACTH Adrenal cortex Cortisol/Corticosterone Components of stress system

4. Tomiyama et al., 2011 Epel et al., 2004 One subset of the population increases food intake under stress and conversely, another decreases food intake. Block et al., 2009

5. Foster et al., 2009; la Fleur et al., 2005 Stress and food preference

6. Access to sucrose reduces the activation of the HPA axis in response to stress. Foster et al., 2009 Consumption of more palatable food improves emotional states, as reflected by reduced anxiety- and depressive-type behaviors. Maniam and Morris, 2010 Ulrich-Lai et al., 2010

7. Foot shock stress – short-term stress (120 electric paw shock; 1 mA, 1 s; 30 min/day; 3 consecutive days). Chronic unpredictable mild stress (CUMS) – 14 days. Cafeteria diet High carbohidrate High lipids Ortolani et al., 2011; 2014 Regular chow + comfort food Stress models used in the Laboratory of Stress Biology

8. light intensity: 60 lux, behavior was registered during 5 min. Analized parameters: number of entries time spent in the open and closed arms number of head dipping, rearing, fecal bolus, stretched-attend posture, risk assessment and grooming. Analized parameters: latency of first crossing, time spent in the periphery and in the center number of crossing, rearing, grooming and fecal bolus. Behavioral analysis EPMEZMOF

9. Ortolani et al., 2011 Activation of autonomic nervous system; Release of CRH, ACTH, glucocorticoids, leptin and insulin. Many factors may be envolved: The mechanisms have not been clarified yet Foot shock stress reduces the intake of regular chow but not that of comfort food

10. Ortolani et al., 2014 CUMS reduces the intake of commercial chow and comfort food

11. Ortolani et al., 2011; 2014 Comfort food intake attenuates endocrine response to foot shock stress and CUMS

12. Ortolani et al., 2011 Foot shock stress reduces anxiety-like behaviors – EPM. This effect is not altered by comfort food.

13. Ortolani et al., 2011 Foot shock stress reduces anxiety-like behaviors – EPM. This effect is not altered by comfort food. + +

14. Ortolani et al., 2011 Foot shock stress associated to comfort reduces anxiety-like behaviors - open field.

15. CUMS (14 days) induces anxiety-like behavior - EPM. This effect is not altered by comfort food. Ortolani et al., 2014

16. CUMS (14 days) induces anxiety-like behavior - open field. This effect is not altered by comfort food. Ortolani et al., 2014

17. Conclusions The access to comfort food attenuates the corticosterone response to stress but did not prevent anxiety-like behaviors of rats exposed to chronic stress.

18. Chronic oral corticosterone was recently reported to induce impressive metabolic changes in mice including weight gain, increased adiposity, elevated plasma leptin, insulin and triglyceride levels, and hyperphagia (Karatsoreos et al., 2010). This model has the added benefit of result in a late-night increase in plasma corticosterone, mimicking one of the most predictive factors in Cushing’s syndrome (Yaneva et al., 2004).0 Sarabdjitsingh et al., 2012

19. HCM system 17.9 ug/day/g corticosterone or vehicle (1% ethanol solution) in the drinking water during a 14 day monitoring period.

20. 20

22. 22 OrexigenicAnorexigenic Co-expressed in arcuate nucleus neurons * mRNA expression of hypothalamic genes

23. 23 Daily food intake Daily liquid intake Results

24. Locomotor activity in the PAS equipment

25. EZM analysis

26. Novel object analysis Blue line: control group Green line: CORT group

27. 27 Red line: control group Blue line: CORT group

28. 28 Bout size Red line: control group Blue line: CORT group

29. Red line: control group Blue line: CORT group

30. Red line: control group Blue line: CORT group

31. Chronic CORT treatment alters components of behavioral patterns and their circadian regulation, and results in a phenotype that mimics the metabolic syndrome. CORT-induced suppression of DC activity is mediated by a selective suppression of AS duration, rather than changes in locomotion bout properties. CORT-induced enhancement of feeding arrives from a selective increase in feeding bout size driven by prolonged feeding bout durations. These findings suggest a deficit in satiation or the process that terminates a meal in CORT-treated mice. Chronic CORT treatment suppressed exploration of a novel object, a finding consistent with an enhancement of anxiety-related behavior. Conclusions

32. Spadari’s Lab – Laboratory of Stress Biology (Federal University of Sao Paulo, Santos, Brazil) Acknowledgments

33. Dr. Laurance Tecott (University of California, San Francisco, CA, USA) Financial Support: Judith Rose Shea Foundation and FAPESP Acknowledgments

34. regina.spadari@unifesp.br Thank you!

35. Chronic unpredictable mild stress (CUMS): 14 days Moreau et al., 1994; Ortolani et al., 2014 PeriodMondayTuesdayWednesdayThursdayFriday Morning Restraint (30 min) Restraint (30 min) Food restriction Empty water bottle (60 min) Restraint (30 min) Restraint (30 min) Afternoon Restraint (30 min) Restraint (30 min) Restraint (30 min) Restraint (30 min) Night Light on during the night Water/food deprivation overnight Water deprivation overnight Damp sawdust overnight Inversion of the light/dark cycle over weekend

36. Comfort food Cafeteria food: commercial chow (Labina, Purina®, Grupo Evialis), peanuts (Hikari®, São Paulo), milk chocolate (Chocolates Garoto®) and biscuit (Tostines®, Nestlé). ComponentsCommercial ChowComfort Food Protein23,5%20% Lipid4,6%20% Carbohidrate33,4%48% Fiber11,4%4% Total Caloric Value17,03 kJ/g21,40 kJ/g Ortolani et al., 2011; 2014