Cannabinoid receptor type 2 (CB2R) plays a critical role in nociception. In contrast to cannabinoid receptor type 1 ligands, CB2R agonists do not produce undesirable central nervous system effects and thus promise to treat neuropathic pain that is often resistant to medical therapy. In the study presented here, we evaluated the bilateral distribution of the CB2R protein and messenger RNA (mRNA) in rat dorsal root ganglia (DRG) after unilateral peripheral nerve injury using immunohistochemistry, western blot, and in situ hybridization analysis. Unilateral chronic constriction injury (CCI) of the sciatic nerve induced neuropathic pain behavior and bilateral elevation of both CB2R protein and mRNA in lumbar L4-L5 as well as cervical C7-C8 DRG when compared with naive animals. CB2R protein and mRNA were increased not only in DRG neurons but also in satellite glial cells. The fact that changes appear bilaterally and (albeit at a lower level) even in the remote cervical DRG can be related to propagation of neuroinflammation alongside the neuraxis and to the neuroprotective effects of CB2R.

Central European Institute of Technology Brno Czech Republic

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Anand U, Otto WR, Sanchez-Herrera D, Facer P, Yiangou Y, Korchev Y, Birch R, Benham C, Bountra C, Chessell IP, et al. 2008. Cannabinoid receptor CB2 localisation and agonist-mediated inhibition of capsaicin responses in human sensory neurons. Pain. 138:667–680 PubMed

Atwood BK, Mackie K. 2010. CB2: a cannabinoid receptor with an identity crisis. Br J Pharmacol. 160:467–479 PubMed PMC

Atwood BK, Straiker A, Mackie K. 2012. CB2 cannabinoid receptors inhibit synaptic transmission when expressed in cultured autaptic neurons. Neuropharmacology. 63:514–523 PubMed PMC

Austin PJ, Moalem-Taylor G. 2010. The neuro-immune balance in neuropathic pain: involvement of inflammatory immune cells, immune-like glial cells and cytokines. J Neuroimmunol. 229:26–50 PubMed

Beltramo M, Bernardini N, Bertorelli R, Campanella M, Nicolussi E, Fredduzzi S, Reggiani A. 2006. CB2 receptor-mediated antihyperalgesia: possible direct involvement of neural mechanisms. J Neurosci. 23:1530–1538 PubMed

Bouaboula M, Desnoyer N, Carayon P, Combes T, Casellas P. 1999. Gi protein modulation induced by a selective inverse agonist for the peripheral cannabinoid receptor CB2: implication for intracellular signalization cross-regulation. Mol Pharmacol. 55:473–480 PubMed

Brázda V, Klusáková I, Svíženská I, Veselková Z, Dubový P. 2009. Bilateral changes in IL-6 protein, but not in its receptor gp130, in rat dorsal root ganglia following sciatic nerve ligature. Cell Mol Neurobiol. 29:1053–1062 PubMed

Brázda V, Muller P, Brozkova K, Vojtesek B. 2006. Restoring wild-type conformation and DNA-binding activity of mutant p53 is insufficient for restoration of transcriptional activity. Biochem Biophys Res Commun. 351:499–506 PubMed

Brooks JW, Thompson SW, Rice AS, Malcanqio M. 2004. (S)-AMPA inhibits electrically evoked calcitonin gene–related peptide (CGRP) release from the rat dorsal horn: reversal by cannabinoid receptor antagonist SR141716A. Neurosci Lett. 372:85–88 PubMed

Brownjohn PW, Ashton JC. 2009. Novel targets in pain research: the case for CB2 receptors as a biorational pain target. Curr Anaesth Crit Care. 20:198–203

Burry RW. 2000. Specificity controls for immunocytochemical methods. J Histochem Cytochem. 48:163–165 PubMed

Calignano A, La Rana G, Giuffrida A, Piomelli D. 1998. Control of pain initiation by endogenous cannabinoids. Nature. 394:277–281 PubMed

Clark AK, Gentry C, Bradbury EJ, McMahon SB, Malcangio M. 2007. Role of spinal microglia in rat models of peripheral nerve injury and inflammation. Eur J Pain. 11:223–230 PubMed

DeLeo JA, Yezierski RP. 2001. The role of neuroinflammation and neuroimmune activation in persistent pain. Pain. 90:1–; 6 PubMed

Devor M, Seltzer Z. 1999. Pathophysiology of damaged nerves in relation to chronic pain. In: Wall PD, Melzack R, editors. Textbook of pain, 4th ed. Edinburgh, UK: Churchill Livingstone; p. 129–164

Dijkstra CD, Damoiseaux J. 1993. Macrophage heterogeneity established by immunocytochemistry. Prog Histochem Cytochem. 27:1–65 PubMed

Dublin P, Hanani M. 2007. Satellite glial cells in sensory ganglia: their possible contribution to inflammatory pain. Brain Behav Immun. 21:592–598 PubMed

Dubový P. 2011. Wallerian degeneration and peripheral nerve conditions for both axonal regeneration and neuropathic pain induction. Ann Anat. 193:267–275 PubMed

Dubový P, Jančálek R, Klusáková I, Svíženská I, Pejchalova K. 2006. Intra- and extraneuronal changes of immunofluorescence staining for TNF-alpha and TNFR1 in the dorsal root ganglia of rat peripheral neuropathic pain models. Cell Mol Neurobiol. 26:1205–1217 PubMed

Dubový P, Klusáková I, Svíženská I. 2002. A quantitative immunohistochemical study of the endoneurium in the rat dorsal and ventral spinal roots. Histochem Cell Biol. 117:473–480 PubMed

Dubový P, Klusáková I, Svíženská I, Brázda V. 2010a. Satellite glial cells express IL-6 and corresponding signal-transducing receptors in the dorsal root ganglia of rat neuropathic pain model. Neuron Glia Biol. 6:73–83 PubMed

Dubový P, Klusáková I, Svíženská I, Brázda V. 2010b. Spatio-temporal changes of SDF1 and its CXCR4 receptor in the dorsal root ganglia following unilateral sciatic nerve injury as a model of neuropathic pain. Histochem Cell Biol. 133:323–337 PubMed

Dubový P, Tuckova L, Jančálek R, Svíženská I, Klusáková I. 2007. Increased invasion of ED-1 positive macrophages in both ipsi- and contralateral dorsal root ganglia following unilateral nerve injuries. Neurosci Lett. 427:88–93 PubMed

Elson K, Simmons A, Speck P. 2004. Satellite cell proliferation in murine sensory ganglia in response to scarification of the skin. Glia. 45:105–109 PubMed

Fernández-Ruiz J, González S, Romero J, Ramos JA. 2005. Cannabinoids in neurodegeneration and neuroprotection. In: Mechoulam R, editor. Cannabinoids as therapeutics. Basel, Switzerland: Birkhäuser Verlag; p. 79–109.10.1007/3-7643-7358-X_5 DOI

Gao Y, Vasilyev DV, Goncalves MB, Howell FV, Hobbs C, Mechoulam R, Shen R, Zhang MY, Strassle BW, Lu P, et al. 2010. Loss of retrograde endocannabinoids signaling and reduced adult neurogenesis in diacylglycerol lipase knock-out mice. J Neurosci. 30:2017–2024 PubMed PMC

Hanani M. 2005. Satellite glial cells in sensory ganglia: from form to function. Brain Res Brain Res Rev. 48:457–476 PubMed

Hanuš L, Breuer A, Tchilibon S, Shiloah S, Goldenberg D, Horowitz M, Pertwee RG, Ross RA, Mechoulam R, Fride E. 1999. HU-308 a specific agonist for CB(2), a peripheral cannabinoid receptor. Proc Natl Acad Sci U S A. 96:14228–14233 PubMed PMC

Hargreaves K, Dubner R, Brown F, Flores C, Joris J. 1988. A new and sensitive method for measuring thermal nociception in cutaneous hyperalgesia. Pain. 32:77–88 PubMed

Harnicarova A, Kozubek S, Pachernik J, Krejci J, Bartova E. 2006. Distinct nuclear arrangement of active and inactive c-myc genes in control and differentiated colon carcinoma cells. Exp Cell Res. 312:4019–4035 PubMed

Hill CE, Harrison BJ, Rau KK, Hougland MT, Bunge MB, Mendell LM, Petruska JC. 2010. Skin incision induces expression of axonal regeneration-related genes in adult rat spinal sensory neurons. J Pain. 11:1066–1073 PubMed PMC

Hohmann AG, Martin WJ, Tsou K, Walker JM. 1995. Inhibition of noxious stimulus-evoked activity of spinal cord dorsal horn neurons by the cannabinoid WIN 55,212-2. Life Sci. 56:2111–2118 PubMed

Hsieh GC, Pai M, Chandran P, Hooker BA, Zhu CZ, Salyers AK, Wensink EJ, Zhan CC, Carroll WA, Dart MJ, et al. 2011. Central and peripheral sites of action for CB2 receptor mediated analgesic activity in chronic inflammatory and neuropathic pain models in rats. Br J Pharmacol. 162:428–440 PubMed PMC

Hu P, Bembrick AL, Keay KA, McLachlan EM. 2007. Immune cell involvement in dorsal root ganglia and spinal cord after chronic constriction or transection of the rat sciatic nerve. Brain Behav Immun. 21:599–616 PubMed

Hu P, McLachlan EM. 2003. Distinct functional types of macrophage in dorsal root ganglia and spinal nerves proximal to sciatic and spinal nerve transections in the rat. Exp Neurol. 184:590–605 PubMed

Huang TY, Cherkas PS, Rosenthal DW, Hanani M. 2005. Dye coupling among satellite glial cells in mammalian dorsal root ganglia. Brain Res. 1036:42–49 PubMed

Ibrahim MM, Deng H, Zvonok A, Cockayne DA, Kwan J, Mata HP, Vanderah TW, Lai J, Porreca F, Makriyannis A, et al. 2003. Activation of CB2 cannabinoid receptors by AM1241 inhibits experimental neuropathic pain: pain inhibition by receptors not present in the CNS. Proc Natl Acad Sci U S A. 100:10529–10533 PubMed PMC

Jacobs JM, Macfarlane RM, Cavanagh JB. 1976. Vascular leakage in the dorsal root ganglia of the rat, studied with horseradish peroxidase. J Neurol Sci. 29:95–107 PubMed

Jančálek R, Dubový P, Svíženská I, Klusáková I. 2010. Bilateral changes of TNFalpha and IL-10 protein in the lumbar and cervical dorsal root ganglia following a unilateral chronic constriction injury of the sciatic nerve. J Neuroinflamm. 7:11 PubMed PMC

Jančálek R, Svíženská HI, Klusáková I, Dubový P. 2011. Bilateral changes of IL-10 protein in lumbar and cervical dorsal root ganglia following proximal and distal chronic constriction injury of peripheral nerve. Neurosci Lett. 501:86–91 PubMed

Jasmin L, Vit J-P, Bhargava A, Ohara PT. 2010. Can satellite glial cells be therapeutic target for pain control? Neuron Glia Biol. 6:63–71 PubMed PMC

Jean-Gilles L, Gran B, Constantinescu CS. 2010. Interaction between cytokines, cannabinoids and the nervous system. Immunobiology. 215:606–610 PubMed

Kirita T, Takebayashi T, Mizuno S, Takeuchi H, Kobayashi T, Fukao M, Yamashita T, Tohse N. 2007. Electrophysiologic changes in dorsal root ganglion neurons and behavioral changes in a lumbar radiculopathy model. Spine. 32:E65–E72 PubMed

Klein TW. 2005. Cannabinoid-based drugs as anti-inflammatory therapeutics. Nat Rev Immunol. 5:400–411 PubMed

Kleinschnitz C, Brinkhoff J, Sommer C, Stoll G. 2005. Contralateral cytokine gene induction after peripheral nerve lesions: dependence on the mode of injury and NMDA receptor signaling. Mol Brain Res. 136:23–28 PubMed

Klusáková I, Dubový P. 2009. Experimental models of peripheral neuropathic pain based on traumatic nerve injuries—an anatomical perspective. Ann Anat. 191:248–259 PubMed

Koltzenburg M, Wall PD, McMahon SB. 1999. Does the right side know what the left is doing? Trends Neurosci. 22:122–127 PubMed

Levin ME, Jin JG, Ji RR, Tong J, Pomonis JD, Lavery DJ, Miller SW, Chiang LW. 2008. Complement activation in the peripheral nervous system following the spinal nerve ligation model of neuropathic pain. Pain. 137:182–201 PubMed

Liu CN, Devor M, Waxman SG, Kocsis JD. 2002. Subthreshold oscillations induced by spinal nerve injury in dissociated muscle and cutaneous afferents of mouse DRG. J Neurophysiol. 87:2009–2017 PubMed PMC

Liu F-Y, Sun Y-N, Wang F-T, Li Q, Su L, Zhao Z-F, Meng X-L, Zhao H, Wu X, Sun Q, et al. 2012. Activation of satellite glial cells in lumbar dorsal root ganglia contributes to neuropathic pain after spinal nerve ligation. Brain Res. 1427:65–77 PubMed

Lu X, Richardson PM. 1991. Inflammation near the nerve cell body enhances axonal regeneration. J. Neurosci. 11:972–978 PubMed PMC

Malan TP, Ibrahim MM, Deng H, Liu Q, Mata HP, Vanderah T, Porreca F, Makriyannis A. 2001. CB2 cannabinoid receptor–mediated peripheral antinociception. Pain. 93:239–245 PubMed

Martin WJ, Hohmann AG, Walker JM. 1996. Suppression of noxious stimulus-evoked activity in the ventral posterolateral nucleus of the thalamus by a cannabinoid agonist: correlation between electrophysiological and antinociceptive effects. J Neurosci. 16:6601–6611 PubMed PMC

Mellick RS, Cavanagh JB. 1968. Changes in blood vessel permeability during degeneration and regeneration in peripheral nerves. Brain. 91:141–160 PubMed

Mitrirattanakul S, Ramakul N, Guerrero AV, Matsuka Y, Ono T, Iwase H, Mackie K, Faull KF, Spigelman I. 2006. Site-specific increases in peripheral cannabinoid receptors and their endogenous ligands in a model of neuropathic pain. Pain. 126:102–114 PubMed PMC

Moalem G, Tracey DJ. 2006. Immune and inflammatory mechanisms in neuropathic pain. Brain Res Rev. 51:240–264 PubMed

Nachemson AK, Bennett GJ. 1993. Does pain damage spinal cord neurons? Transsynaptic degeneration in rat following a surgical incision. Neurosci Lett. 162:78–80 PubMed

Ohara PT, Vit JP, Bhargava A, Romero M, Sundberg C, Charles AC, Jasmin L. 2009. Gliopathic pain: when satellite glial cells go bad. Neuroscientist. 15:450–463 PubMed PMC

Pertwee RG. 2001. Cannabinoid receptors and pain. Prog Neurobiol. 63:569–611 PubMed

Pitcher GM, Ritchie J, Henry LJ. 1999. Nerve constriction in the rat: model of neuropathic, surgical and central pain. Pain. 83:37–46 PubMed

Racz I, Nadal X, Alferink J, Baños JE, Rehnelt J, Martín M, Pintado B, Gutierrez-Adan A, Sanguino E, Manzanares J, et al. 2008. Crucial role of CB(2) cannabinoid receptor in the regulation of central immune responses during neuropathic pain. J Neurosci. 28:12125–12135 PubMed PMC

Richardson JD, Kilo S, Hargreaves KM. 1998. Cannabinoids reduce hyperalgesia and inflammation via interaction with peripheral CB1 receptors. Pain. 75:111–119 PubMed

Ross RA, Coutts AA, McFarlane SM, Anavi-Goffer S, Irving AJ, Pertwee RG, MacEwan DJ, Scott RH. 2001. Actions of cannabinoid receptor ligands on rat cultured sensory neurones: implications for antinociception. Neuropharmacology. 40:221–232 PubMed

Rotshenker S. 2011. Wallerian degeneration: the innate immune response to traumatic nerve injury. J Neuroinflammation. 8:109. PubMed PMC

Ruohonen S, Khademi M, Jagodic M, Taskinen H-S, Olsson T, Roytta M. 2005. Cytokine responses during chronic denervation. J Neuroinflamm. 2:26 PubMed PMC

Sagar DR, Kelly S, Millns PJ, O’Shaughnessey CT, Kendall DA, Chapman V. 2005. Inhibitory effects of CB1 receptor and CB2 receptor agonists on responses of DRG neurons and dorsal horn neurons in neuropathic rats. Eur J Neurosci. 22:371–379 PubMed

Saňudo-Peňa MC, Romero J, Seale GE, Fernandez-Ruiz JJ, Walker JM. 2000. Activational role of cannabinoids on movement. Eur J Pharmacol. 391:269–274 PubMed

Scholz J, Woolf CJ. 2007. The neuropathic pain triad: neurons, immune cells and glia. Nat Neurosci. 10:1361–1368 PubMed

Scott DA, Wright CE, Angus JA. 2004. Evidence that CB-1 and CB-2 cannabinoid receptors mediate antinociception in neuropathic pain in the rat. Pain. 109:124–131 PubMed

Ständer S, Schmelz M, Metze D, Luger T, Rukwied R. 2005. Distribution of cannabinoid receptor 1 (CB1) and 2 (CB2) on sensory nerve fibers and adnexal structures in human skin. J Dermatol Sci. 38:177–188 PubMed

Taiwo YO, Bjerknes LK, Goetzl EJ, Levine JD. 1989. Mediation of primary afferent peripheral hyperalgesia by the cAMP second messenger system. Neuroscience. 32:577–580 PubMed

Taiwo YO, Levine JD. 1991. Further confirmation of the role of adenyl cyclase and of cAMP-dependent protein kinase in primary afferent hyperalgesia. Neuroscience. 44:131–135 PubMed

Takeda K, Muramatsu M, Chikuma T, Kato T. 2009. Effect of memantine on the levels of neuropeptides and microglial cells in the brain regions of rats with neuropathic pain. J Mol Neurosci. 39:380–390 PubMed

Takeda M, Takahashi M, Matsumoto S. 2009. Contribution of the activation of satellite glia in sensory ganglia to pathological pain. Neurosci Biobehav Rev. 33:784–792 PubMed

Takeda M, Tanimoto T, Kadoi J, Nasu M, Takahashi M, Kitagawa J, Matsumoto S. 2007. Enhanced excitability of nociceptive trigeminal ganglion neurons by satellite glial cytokine following peripheral inflammation. Pain 129:155–166 PubMed

Tanimura A, Yamazaki M, Hashimotodani Y, Uchigashima M, Kawata S, Abe M, Kita Y, Hashimoto K, Shimizu T, Watanabe M, et al. 2010. The endocannabinoid 2-arachidonoylglycerol produced by diacylglycerol lipase alpha mediates retrograde suppression of synaptic transmission. Neuron. 65:320–327 PubMed

Tsou K, Lowitz KA, Hohmann AG, Martin WJ, Hathaway CB, Bereiter DA, Walker JM. 1996. Suppression of noxious stimulus-evoked expression of FOS protein-like immunoreactivity in rat spinal cord by a selective cannabinoid agonist. Neuroscience. 70:791–798 PubMed

Üçeyler N, Sommer C. 2006. Wallerian degeneration and neuropathic pain. Drug Discov Today Dis Mech. 3:351–356

Viscomi MT, Oddi S, Latini L, Bisicchia E, Maccarrone M, Molinari M. 2010. The endocannabinoid system: a new entry in remote cell death mechanisms. Exp Neurol. 224:56–65 PubMed

Viscomi MT, Oddi S, Latini L, Pasquariello N, Florenzano F, Bernardi G, Molinari M, Maccarrone M. 2009. Selective CB2 receptor agonism protects central neurons from remote axotomy-induced apoptosis through the PI3 K/Akt pathway. J Neurosci. 29:4564–4570 PubMed PMC

Walczak J-S, Pichette V, Leblond F, Desbiens K, Beaulieu P. 2005. Behavioural, pharmacological and molecular characterization of the saphenous nerve partial ligation: a new model of neuropathic pain. Neuroscience. 132:1093–1102 PubMed

Whiteside GT, Lee GP, Valenzano KJ. 2007. The role of the cannabinoid CB2 receptor in pain transmission and therapeutic potential of small molecule CB2 receptor agonists. Curr Med Chem. 14:917–936 PubMed

Wilkerson JL, Gentry KR, Dengler EC, Wallace JA, Kerwin AA, Kuhn MN, Zvonok AM, Thakur GA, Makriyannis A, Milligan ED. 2012. Immunofluorescent spectral analysis reveals the intrathecal cannabinoid agonist, AM1241, produces spinal anti-inflammatory cytokine responses in neuropathic rats exhibiting relief from allodynia. Brain Behav. 2:155–177 PubMed PMC

Woolf CJ, Mannion RJ. 1999. Neuropathic pain: aetiology, symptoms, mechanisms, and management. Lancet. 353:1959–1964 PubMed

Wotherspoon G, Fox A, McIntyre P, Colley S, Bevan S, Winter J. 2005. Peripheral nerve injury induces cannabinoid receptor 2 protein expression in rat sensory neurons. Neuroscience. 135:235–245 PubMed

Yamaguchi H, Ochi M, Mori R, Ryoke K, Yamamoto S, Iwata A, Uchio Y. 1999. Unilateral sciatic nerve injury stimulates contralateral nerve regeneration. Neuroreport. 10:1359–1362 PubMed

Zamboni L, De Martino C. 1967. Buffered picric acid–formaldehyde—a new rapid fixative for electron microscopy. J Cell Biol. 35:A148

Zhang J, Hoffert C, Vu HK, Groblewski T, Ahmad S, O’Donnel D. 2003. Induction of CB2 receptor expression in the rat spinal cord neuropathic but not inflammatory chronic pain models. Eur J Neurosci. 17:2750–2754 PubMed

Dynamic response to peripheral nerve injury detected by in situ hybridization of IL-6 and its receptor mRNAs in the dorsal root ganglia is not strictly correlated with signs of neuropathic pain