Central Role of Protein Kinase A in Promoting Trigeminal Nociception in an In Vivo Model of Temporomandibular Disorders

Aims: To investigate cellular changes in the spinal trigeminal nucleus (STN) and trigeminal ganglion (TG) associated with trigeminal nociception mediated by inflammation in the temporomandibular joint (TMJ). Methods: Male Sprague-Dawley rats (n = 86) were utilized to investigate cellular and behavioral responses to prolonged TMJ inflammation caused by bilateral injection of Complete Freund’s Adjuvant (CFA) in the TMJ capsules. To investigate the cellular effects of protein kinase A (PKA) in the STN, rats were injected intrathecally with the selective PKA inhibitor KT5720 prior to injection of CFA into both TMJ capsules. Levels of calcitonin gene-related peptide (CGRP), active PKA, and ionized calcium-binding adapter molecule 1 (Iba1) in the STN and expression of phosphorylated extracellular regulated kinases (p-ERK) in the TG were determined with immunohistochemistry (n ≥ 3 experiments per test condition). Nocifensive head withdrawal responses to mechanical stimulation of the cutaneous tissue over the TMJ were monitored following CFA injection in the absence or presence of KT5720 (n = 7). Statistical analysis was performed using parametric analysis of variance (ANOVA) tests. Results: Intrathecal injection of KT5720 significantly inhibited the stimulatory effect of CFA on levels of CGRP, PKA, and Iba1 in the STN. In addition, administration of KT5720 decreased the average number of CFA-induced nocifensive withdrawal responses to mechanical stimulation and the CFA-mediated increase in p-ERK expression in the ganglion. Conclusion: These findings provide evidence that elevated PKA activity in the STN promotes cellular events temporally associated with trigeminal nociception caused by prolonged TMJ inflammation.

calcitonin gene-related peptide; nociception; protein kinase A; TMJ; trigeminal ganglion

1. Ohrbach R, Fillingim RB, Mulkey F, et al. Clinical findings and pain symptoms as potential risk factors for chronic TMD: Descriptive data and empirically identified domains from the OPPERA case-control study. J Pain 2011;12(suppl):t27–t45.

2. Furquim BD, Flamengui LM, Conti PC. TMD and chronic pain: A current view. Dental Press J Orthod 2015;20:127–133.

3. Greenspan JD, Slade GD, Bair E, et al. Pain sensitivity risk factors for chronic TMD: Descriptive data and empirically identified domains from the OPPERA case control study. J Pain 2011;12(suppl):t61–t74.

4. Poveda Roda R, Bagan JV, Díaz Fernández JM, Hernández Bazán S, Jiménez Soriano Y. Review of temporomandibular joint pathology. Part I: Classification, epidemiology and risk factors. Med Oral Patol Oral Cir Bucal 2007;12:e292–e298.

5. Bonjardim LR, Lopes-Filho RJ, Amado G, Albuquerque RL Jr, Goncalves SR. Association between symptoms of temporomandibular disorders and gender, morphological occlusion, and psychological factors in a group of university students. Indian J Dent Res 2009;20:190–194.

6. Maixner W, Fillingim R, Booker D, Sigurdsson A. Sensitivity of patients with painful temporomandibular disorders to experimentally evoked pain. Pain 1995;63:341–351.

7. Maixner W, Fillingim R, Sigurdsson A, Kincaid S, Silva S. Sensitivity of patients with painful temporomandibular disorders toexperimentally evoked pain: Evidence for altered temporal summation of pain. Pain 1998;76:71–81.

8. Bereiter DA, Okamoto K, Bereiter DF. Effect of persistent monoarthritis of the temporomandibular joint region on acute mustard oil-induced excitation of trigeminal subnucleus caudalis neurons in male and female rats. Pain 2005;117:58–67.

9. Shimizu K, Guo W, Wang H, et al. Differential involvement of trigeminal transition zone and laminated subnucleus caudalisin orofacial deep and cutaneous hyperalgesia: The effects of interleukin-10 and glial inhibitors. Mol Pain 2009;5:75.

10. Sessle BJ. Peripheral and central mechanisms of orofacial inflammatory pain. Int Rev Neurobiol 2011;97:179–206.

11. Seybold VS. The role of peptides in central sensitization. Handb Exp Pharmacol 2009;(194):451–491.

12. Durham PL, Garrett FG. Emerging importance of neuron-satellite glia interactions within trigeminal ganglia in craniofacial pain. TOPAINJ 2010;3:3–13.

13. Xie YF. Glial involvement in trigeminal central sensitization. Acta Pharmacol Sin 2008;29:641–645.

14. Davies AJ, Kim YH, Oh SB. Painful neuron-microglia interactions in the trigeminal sensory system. TOPAINJ 2010:14–28.

15. Ikeda H, Kiritoshi T, Murase K. Contribution of microglia and astrocytes to the central sensitization, inflammatory and neuropathic pain in the juvenile rat. Mol Pain 2012;8:43.

16. Moreno MJ, Terrón JA, Stanimirovic DB, Doods H, Hamel E. Characterization of calcitonin gene-related peptide (CGRP) receptors and their receptor-activity-modifying proteins (RAMPs) in human brain microvascular and astroglial cells in culture. Neuropharmacology 2002;42:270–280.

17. Marvizón JC, Pérez OA, Song B, et al. Calcitonin receptor-like receptor and receptor activity modifying protein 1 in the rat dorsal horn: Localization in glutamatergic presynaptic terminals containing opioids and adrenergic alpha2C receptors. Neuroscience 2007;148:250–265.

18. Lennerz JK, Rühle V, Ceppa EP, et al. Calcitonin receptor-like receptor (CLR), receptor activity-modifying protein 1 (RAMP1), and calcitonin gene-related peptide (CGRP) immunoreactivity in the rat trigeminovascular system: Differences between peripheral and central CGRP receptor distribution. J Comp Neurol 2008;507:1277–1299.

19. Hong Y, Hay DL, Quirion R, Poyner DR. The pharmacology of adrenomedullin 2/intermedin. Br J Pharmacol 2012;166:110–120.

20. Russell FA, King R, Smillie SJ, Kodji X, Brain SD. Calcitonin gene-related peptide: Physiology and pathophysiology. Physiol Rev 2014;94:1099–1142.

21. Bao Y, Jiang L, Chen H, Zou J, Liu Z, Shi Y. The neuroprotective effect of liraglutide is mediated by glucagon-like peptide 1 receptor-mediated activation of cAMP/PKA/CREB pathway. Cell Physiol Biochem 2015;36:2366–2378.

22. Waltereit R, Weller M. Signaling from cAMP/PKA to MAPK and synaptic plasticity. Mol Neurobiol 2003;27:99–106.

23. Staud R. Cytokine and immune system abnormalities in fibromyalgia and other central sensitivity syndromes. Curr Rheumatol Rev 2015;11:109–115.

24. Sun RQ, Tu YJ, Lawand NB, Yan JY, Lin Q, Willis WD. Calcitonin gene-related peptide receptor activation produces PKA- and PKC-dependent mechanical hyperalgesia and central sensitization. J Neurophysiol 2004;92:2859–2866.

25. Thalakoti S, Patil VV, Damodaram S, et al. Neuron-glia signaling in trigeminal ganglion: Implications for migraine pathology. Headache 2007;47:1008–1023.

26. Cady RJ, Glenn JR, Smith KM, Durham PL. Calcitonin gene-related peptide promotes cellular changes in trigeminal neurons and glia implicated in peripheral and central sensitization. Mol Pain 2011;7:94.

27. Ji RR, Gereau RW 4th, Malcangio M, Strichartz GR. MAP kinase and pain. Brain Res Rev 2009;60:135–148.

28. Cady RJ, Denson JE, Sullivan LQ, Durham PL. Dual orexin receptor antagonist 12 inhibits expression of proteins in neurons and glia implicated in peripheral and central sensitization. Neuroscience 2014;269:79–92.

29. Villa G, Ceruti S, Zanardelli M, et al. Temporomandibular joint inflammation activates glial and immune cells in both the trigeminal ganglia and in the spinal trigeminal nucleus. Mol Pain 2010;6:89.

30. Yamazaki Y, Ren K, Shimada M, Iwata K. Modulation of paratrigeminal nociceptive neurons following temporomandibular joint inflammation in rats. Exp Neurol 2008;214:209–218.

31. Hawkins JL, Denson JE, Miley DR, Durham PL. Nicotine stimulates expression of proteins implicated in peripheral and central sensitization. Neuroscience 2015;290:115–125.

32. Garrett FG, Hawkins JL, Overmyer AE, Hayden JB, Durham PL. Validation of a novel rat-holding device for studying heat- and mechanical-evoked trigeminal nocifensive behavioral responses. J Orofac Pain 2012;26:337–344.

33. Hawkins JL, Durham PL. Prolonged jaw opening promotes nociception and enhanced cytokine expression. J Oral Facial Pain Headache 2016;30:34–41.

34. Harper RP, Kerins CA, McIntosh JE, Spears R, Bellinger LL. Modulation of the inflammatory response in the rat TMJ with increasing doses of complete Freund’s adjuvant. Osteoarthritis Cartilage 2001;9:619–624.

35. Imbe H, Iwata K, Zhou QQ, Zou S, Dubner R, Ren K. Orofacial deep and cutaneous tissue inflammation and trigeminal neuronal activation. Implications for persistent temporomandibular pain. Cells Tissues Organs 2001;169:238–247.

36. Sato T, Kitagawa J, Ren K, et al. Activation of trigeminal intranuclear pathway in rats with temporomandibular joint inflammation. J Oral Sci 2005;47:65–69.

37. Spears R, Dees LA, Sapozhnikov M, Bellinger LL, Hutchins B. Temporal changes in inflammatory mediator concentrations inan adjuvant model of temporomandibular joint inflammation. J Orofac Pain 2005;19:34–40.

38. Romero-Reyes M, Pardi V, Akerman S. A potent and selective calcitonin gene-related peptide (CGRP) receptor antagonist, MK-8825, inhibits responses to nociceptive trigeminal activation: Role of CGRP in orofacial pain. Exp Neurol 2015;271:95–103.

39. Hutchins B, Patel H, Spears R. Attenuation of pro-inflammatory neuropeptide levels produced by a cyclooxygenase-2 inhibitor in an animal model of chronic temporomandibular joint inflammation. J Orofac Pain 2002;16:312–316.

40. Bird GC, Han JS, Fu Y, Adwanikar H, Willis WD, Neugebauer

V. Pain-related synaptic plasticity in spinal dorsal horn neurons: Role of CGRP. Mol Pain 2006;2:31.

41. Sun RQ, Lawand NB, Lin Q, Willis WD. Role of calcitonin gene-related peptide in the sensitization of dorsal horn neurons to mechanical stimulation after intradermal injection of capsaicin. J Neurophysiol 2004;92:320–326.

42. Walker CS, Hay DL. CGRP in the trigeminovascular system: A role for CGRP, adrenomedullin and amylin receptors? Br J Pharmacol 2013;170:1293–1307.

43. Hoffman MS, Mitchell GS. Spinal 5-HT7 receptors and protein kinase A constrain intermittent hypoxia-induced phrenic longterm facilitation. Neuroscience 2013;250:632–643.

44. Fu Y, Han J, Ishola T, et al. PKA and ERK, but not PKC, in the amygdala contribute to pain-related synaptic plasticity and behavior. Mol Pain 2008;4:26.

45. Kohno T, Wang H, Amaya F, et al. Bradykinin enhances AMPA and NMDA receptor activity in spinal cord dorsal horn neurons by activating multiple kinases to produce pain hypersensitivity. J Neurosci 2008;28:4533–4540.

46. Hu HJ, Glauner KS, Gereau RW 4th. ERK integrates PKA and PKC signaling in superficial dorsal horn neurons. I. Modulation of A-type K+ currents. J Neurophysiol 2003;90:1671–1679.

47. Levy D, Strassman AM. Distinct sensitizing effects of the cAMP-PKA second messenger cascade on rat dural mechanonociceptors. J Physiol 2002;538:483–493.

48. Gosselin RD, Suter MR, Ji RR, Decosterd I. Glial cells and chronic pain. Neuroscientist 2010;16:519–531.

49. Ji RR, Berta T, Nedergaard M. Glia and pain: Is chronic pain a gliopathy? Pain 2013;154(suppl):s10–s28.

50. Chiu HY, Lin HH, Lai CC. Potentiation of spinal NMDA-mediated nociception by cocaine- and amphetamine-regulated transcript peptide via PKA and PKC signaling pathways in rats. Regul Pept 2009;158:77–85.

51. Alter BJ, Zhao C, Karim F, Landreth GE, Gereau RW 4th. Genetic targeting of ERK1 suggests a predominant role for ERK2 in murine pain models. J Neurosci 2010;30:11537–11547.

52. Durham PL, Russo AF. Serotonergic repression of mitogen-activated protein kinase control of the calcitonin gene-related peptide enhancer. Mol Endocrinol 1998;12:1002–1009.

53. Wang Z, Ma W, Chabot JG, Quirion R. Cell-type specific activation of p38 and ERK mediates calcitonin gene-related peptide involvement in tolerance to morphine-induced analgesia. FASEB J 2009;23:2576–2586.

54. Cornelison LE, Hawkins JL, Durham PL. Elevated levels of calcitonin gene-related peptide in upper spinal cord promotes sensitization of primary trigeminal nociceptive neurons. Neuroscience 2016;339:491–501.

55. Bagis B, Ayaz EA, Turgut S, Durkan R, Özcan M. Gender difference in prevalence of signs and symptoms of temporomandibular joint disorders: A retrospective study on 243 consecutive patients. Int J Med Sci 2012;9:539–544.

56. Slade GD, Bair E, Greenspan JD, et al. Signs and symptoms of first-onset TMD and sociodemographic predictors of its development: The OPPERA prospective cohort study. J Pain 2013;14(suppl):t20–t32, e1–e3.