2. Effects of monoamine uptake inhibitors in an assay of pain-related depression of behavior in male mice Khadijah Alexander P.I. Dr. Laurence Miller Department of Psychological Sciences

3. Preview Rationale/hypothesis Methods Results Discussion

4. Introduction Pain-depressed behavior is a clinically relevant component of pain (Brennan et al. 2007) Preclinical studies of pain have often ignored this component (Negus et al. 2006) This inconsistency may limit the development of new medications Pain Behavior (e.g. reflexive withdrawal) Pain Behavior (e.g. capacity for work or exercise)

5. Focus of our studies Evidence of a role for dopamine in pain depressed behavior Dopamine –neurotransmitter important in motivated behavior (Floresco 2015) Pain = decrease in dopamine at the nucleus accumbens (Miller et al 2015) Pain Dopamine

6. Hypothesis Prediction: Drugs that increase dopamine activity will block pain-depressed behavior Pain Dopamine Behavior

7. Methods Pharmacological approach Monoamine Uptake Inhibitors

8. Methods Pharmacological approach Monoamine Uptake Inhibitors Monoamine Transporter Monoamine Neurotransmitter Receptor Presynaptic Neuron Postsynaptic Neuron

9. Methods Pharmacological approach Monoamine Uptake Inhibitors Monoamine Uptake Inhibitor Monoamine Neurotransmitter Receptor Postsynaptic Neuron

10. Methods Pharmacological approach Monoamine Uptake Inhibitors Monoamine Uptake Inhibitor Monoamine Neurotransmitter Receptor Presynaptic Neuron Postsynaptic Neuron

11. Methods Pharmacological approach Monoamine Uptake Inhibitors Citalopram (Serotonin) Nisoxetine (Norepinephrine) Milnacipran (Serotonin/Norepinephrine) Buprorpion (Dopamine) Monoamine Uptake Inhibitor Monoamine Neurotransmitter Receptor Presynaptic Neuron Postsynaptic Neuron

12. Methods Behavioral Approach Pain stimulus: Intraperitoneal dilute lactic acid (0.56%)

13. Methods Behavioral Approach Pain stimulus: Intraperitoneal dilute lactic acid (0.56%) Pain-stimulated behavior Acid-stimulated stretching

14. Methods Behavioral Approach Pain stimulus: Intraperitoneal dilute lactic acid (0.56%) Pain-stimulated behavior Acid-stimulated stretching

15. Methods Behavioral Approach Pain stimulus: Intraperitoneal dilute lactic acid (0.56%) Pain-stimulated behavior Acid-stimulated stretching Pain-depressed behavior Acid-depressed nesting

16. Methods Behavioral Approach Pain stimulus: Intraperitoneal dilute lactic acid (0.56%) Pain-stimulated behaviorPain-depressed behavior

17. Methods Behavioral Approach Pain stimulus: Intraperitoneal dilute lactic acid (0.56%) Pain-stimulated behaviorPain-depressed behavior

18. Methods Behavioral Approach Pain stimulus: Intraperitoneal dilute lactic acid (0.56%) Pain-stimulated behaviorPain-depressed behavior

19. Methods Behavioral Approach Pain stimulus: Intraperitoneal dilute lactic acid (0.56%) Pain-stimulated behaviorPain-depressed behavior

20. Methods Behavioral Approach Pain stimulus: Intraperitoneal dilute lactic acid (0.56%) Pain-stimulated behaviorPain-depressed behavior 1 2 3 4 5 Dependent Variable = Number of zones cleared

21. Methods Behavioral Approach Pain stimulus: Intraperitoneal dilute lactic acid (0.56%) Pain-stimulated behaviorPain-depressed behavior *

22. Methods Behavioral Approach Pain stimulus: Intraperitoneal dilute lactic acid (0.56%) Pain-stimulated behaviorPain-depressed behavior *

23. Results Citalopram – Serotonin Selective Uptake Inhibitor Pain-stimulated behavior Pain-depressed behavior filled symbols indicate significant difference compared to control

24. Results Citalopram – Serotonin Selective Uptake Inhibitor Pain-stimulated behavior Pain-depressed behavior filled symbols indicate significant difference compared to control

25. Results Citalopram – Serotonin Selective Uptake Inhibitor Pain-stimulated behavior Pain-depressed behavior filled symbols indicate significant difference compared to control

26. Results Citalopram – Serotonin Selective Uptake Inhibitor Pain-stimulated behavior Pain-depressed behavior filled symbols indicate significant difference compared to control

27. Results Nisoxetine – Norepinephrine Selective Uptake Inhibitor Pain-stimulated behavior Pain-depressed behavior filled symbols indicate significant difference compared to control

28. Results Nisoxetine – Norepinephrine Selective Uptake Inhibitor Pain-stimulated behavior Pain-depressed behavior filled symbols indicate significant difference compared to control

29. Results Nisoxetine – Norepinephrine Selective Uptake Inhibitor Pain-stimulated behavior Pain-depressed behavior filled symbols indicate significant difference compared to control

30. Results Milnacipran – Serotonin+Norepinephrine Uptake Inhibitor Pain-stimulated behavior Pain-depressed behavior filled symbols indicate significant difference compared to control

31. Results Milnacipran – Serotonin+Norepinephrine Uptake Inhibitor Pain-stimulated behavior Pain-depressed behavior filled symbols indicate significant difference compared to control

32. Results Milnacipran – Serotonin+Norepinephrine Uptake Inhibitor Pain-stimulated behavior Pain-depressed behavior filled symbols indicate significant difference compared to control

33. Results Bupropion – Dopamine Selective Uptake Inhibitor Pain-stimulated behavior Pain-depressed behavior filled symbols indicate significant difference compared to control

34. Results Bupropion – Dopamine Selective Uptake Inhibitor Pain-stimulated behavior Pain-depressed behavior filled symbols indicate significant difference compared to control

35. Results Bupropion – Dopamine Selective Uptake Inhibitor Pain-stimulated behavior Pain-depressed behavior filled symbols indicate significant difference compared to control

36. Results Summary DrugPain-Stimulated Behavior Pain-Depressed Behavior KetoprofenClinically effective NSAID CitalopramSerotonin × NisoxetineNorepinephrine × MilnacipranSerotonin+Norepinephrine × BupropionDopamine ?

37. Discussion Results with ketoprofen support the validity of assays of pain- depressed behavior as a complement to traditional approaches Results with monoamine uptake inhibitors lacking effects on dopamine suggest these drugs have limited potential for the treatment of acute pain-related depression of behavior Preliminary results with bupropion indicate that it blocks pain- stimulated behavior, but experiments on pain-depressed behavior are ongoing

38. Acknowledgements Dena Phillips Taylor Rodriguez Amma Sarfo John Shallcross Dr. Tadd Patton Center for Undergraduate Research and Scholarship College of Science and Mathematics Department of Psychological Sciences Office of Faculty Development and Teaching Excellence

39. Questions?

41. Drugs Used in Study Ketoprofen Citalopram (Celexa) Nisoxetine Milnacinpran (Savella, Dalcipran) Bupropion (Wellbutrin)

42. Dorsal Root Ganglion Spinal Cord Thalamus Anterior Cingulate Cortex Rostral Ventromedial Medulla Parabrachial Nucleus Periaqueductal Gray Hypothalamus Insula Somatosensory Cortex II Somatosensory Cortex I

43. Dorsal Root Ganglion Spinal Cord Thalamus Anterior Cingulate Cortex Nucleus Accumbens Ventral Tegmental Area Rostral Ventromedial Medulla Parabrachial Nucleus Periaqueductal Gray Hypothalamus Insula Somatosensory Cortex II Somatosensory Cortex I