Adenoviral-Mediated Glial Cell Line–Derived Neurotrophic Factor Gene Transfer Has a Protective Effect on Sciatic Nerve Following Constriction-Induced Spinal Cord Injury
Neuropathic pain due to peripheral nerve injury may be associated with abnormal central nerve activity. Glial cell-line-derived neurotrophic factor (GDNF) can help attenuate neuropathic pain in different animal models of nerve injury. However, whether GDNF can ameliorate neuropathic pain in the spinal cord dorsal horn (SCDH) in constriction-induced peripheral nerve injury remains unknown. We investigated the therapeutic effects of adenoviral-mediated GDNF on neuropathic pain behaviors, microglial activation, pro-inflammatory cytokine expression and programmed cell death in a chronic constriction injury (CCI) nerve injury animal model. In this study, neuropathic pain was produced by CCI on the ipsilateral SCDH. Mechanical allodynia was examined with von Frey filaments and thermal sensitivity was tested using a plantar test apparatus post-operatively. Target proteins GDNF-1, GDNFRa-1, MMP2, MMP9, p38, phospho-p38, ED1, IL6, IL1β, AIF, caspase-9, cleaved caspase-9, caspase-3, cleaved caspase-3, PARP, cleaved PARP, SPECTRIN, cleaved SPECTRIN, Beclin-1, PKCσ, PKCγ, iNOS, eNOS and nNOS were detected. Microglial activity was measured by observing changes in immunoreactivity with OX-42. NeuN and TUNEL staining were used to reveal whether apoptosis was attenuated by GDNF. Results showed that administrating GDNF began to attenuate both allodynia and thermal hyperalgesia at day 7. CCI-rats were found to have lower GDNF and GDNFRa-1 expression compared to controls, and GDNF re-activated their expression. Also, GDNF significantly down-regulated CCI-induced protein expression except for MMP2, eNOS and nNOS, indicating that the protective action of GDNF might be associated with anti-inflammation and prohibition of microglia activation. Immunocytochemistry staining showed that GDNF reduced CCI-induced neuronal apoptosis. In sum, GDNF enhanced the neurotrophic effect by inhibiting microglia activation and cytokine production via p38 and PKC signaling. GDNF could be a good therapeutic tool to attenuate programmed cell death, including apoptosis and autophagy, consequent to CCI-induced peripheral nerve injury.
References
[1]
Lekan HA, Carlton SM, Coggeshall RE (1996) Sprouting of A beta fibers into lamina II of the rat dorsal horn in peripheral neuropathy. Neurosci Lett 208: 147–150. doi: 10.1016/0304-3940(96)12566-0
[2]
Woolf CJ, Shortland P, Coggeshall RE (1992) Peripheral nerve injury triggers central sprouting of myelinated afferents. Nature 355: 75–78. doi: 10.1038/355075a0
[3]
Shortland P, Wang HF, Molander C (1999) Distribution of transganglionically labelled soybean agglutinin primary afferent fibres after nerve injury. Brain Res 815: 206–212. doi: 10.1016/s0006-8993(98)01152-4
[4]
Bridges D, Thompson SW, Rice AS (2001) Mechanisms of neuropathic pain. Br J Anaesth 87: 12–26. doi: 10.1093/bja/87.1.12
[5]
Airaksinen MS, Saarma M (2002) The GDNF family: signalling, biological functions and therapeutic value. Nat Rev Neurosci 3: 383–394. doi: 10.1038/nrn812
[6]
Bennett DL, Michael GJ, Ramachandran N, Munson JB, Averill S, et al. (1998) A distinct subgroup of small DRG cells express GDNF receptor components and GDNF is protective for these neurons after nerve injury. J Neurosci 18: 3059–3072.
[7]
Boucher TJ, Okuse K, Bennett DL, Munson JB, Wood JN, et al. (2000) Potent analgesic effects of GDNF in neuropathic pain states. Science 290: 124–127. doi: 10.1126/science.290.5489.124
[8]
Hao S, Mata M, Wolfe D, Huang S, Glorioso JC, et al. (2003) HSV-mediated gene transfer of the glial cell-derived neurotrophic factor provides an antiallodynic effect on neuropathic pain. Mol Ther 8: 367–375. doi: 10.1016/s1525-0016(03)00185-0
[9]
Ramer MS, Priestley JV, McMahon SB (2000) Functional regeneration of sensory axons into the adult spinal cord. Nature 403: 312–316. doi: 10.1038/35002084
[10]
Eslamboli A (2005) Assessment of GDNF in primate models of Parkinson's disease: comparison with human studies. Rev Neurosci 16: 303–310. doi: 10.1515/revneuro.2005.16.4.303
[11]
Gill SS, Patel NK, Hotton GR, O'Sullivan K, McCarter R, et al. (2003) Direct brain infusion of glial cell line-derived neurotrophic factor in Parkinson disease. Nat Med 9: 589–595. doi: 10.1038/nm850
[12]
Manabe Y, Nagano I, Gazi MS, Murakami T, Shiote M, et al. (2002) Adenovirus-mediated gene transfer of glial cell line-derived neurotrophic factor prevents motor neuron loss of transgenic model mice for amyotrophic lateral sclerosis. Apoptosis 7: 329–334. doi: 10.1179/016164103101201193
[13]
Wang LJ, Lu YY, Muramatsu S, Ikeguchi K, Fujimoto K, et al. (2002) Neuroprotective effects of glial cell line-derived neurotrophic factor mediated by an adeno-associated virus vector in a transgenic animal model of amyotrophic lateral sclerosis. J Neurosci 22: 6920–6928.
[14]
Bennett GJ, Xie YK (1988) A peripheral mononeuropathy in rat that produces disorders of pain sensation like those seen in man. Pain 33: 87–107. doi: 10.1016/0304-3959(88)90209-6
[15]
Tai MH, Cheng H, Wu JP, Liu YL, Lin PR, et al. (2003) Gene transfer of glial cell line-derived neurotrophic factor promotes functional recovery following spinal cord contusion. Exp Neurol 183: 508–515. doi: 10.1016/s0014-4886(03)00130-4
[16]
Giraud C, Winocour E, Berns KI (1995) Recombinant junctions formed by site-specific integration of adeno-associated virus into an episome. J Virol 69: 6917–6924.
[17]
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. doi: 10.1016/0304-3959(88)90026-7
[18]
Eftekhari S, Edvinsson L (2011) Calcitonin gene-related peptide (CGRP) and its receptor components in human and rat spinal trigeminal nucleus and spinal cord at C1-level. BMC Neurosci 12: 112. doi: 10.1186/1471-2202-12-112
[19]
Yamazaki K, Wakasugi N, Tomita T, Kikuchi T, Mukoyama M, et al. (1988) Gracile axonal dystrophy (GAD), a new neurological mutant in the mouse. Proc Soc Exp Biol Med 187: 209–215. doi: 10.3181/00379727-187-42656
[20]
Kawasaki Y, Xu ZZ, Wang X, Park JY, Zhuang ZY, et al. (2008) Distinct roles of matrix metalloproteases in the early- and late-phase development of neuropathic pain. Nat Med 14: 331–336. doi: 10.1038/nm1723
[21]
Jin SX, Zhuang ZY, Woolf CJ, Ji RR (2003) p38 mitogen-activated protein kinase is activated after a spinal nerve ligation in spinal cord microglia and dorsal root ganglion neurons and contributes to the generation of neuropathic pain. J Neurosci 23: 4017–4022.
[22]
George A, Buehl A, Sommer C (2004) Wallerian degeneration after crush injury of rat sciatic nerve increases endo- and epineurial tumor necrosis factor-alpha protein. Neurosci Lett 372: 215–219. doi: 10.1016/j.neulet.2004.09.075
[23]
Mao J, Price DD, Phillips LL, Lu J, Mayer DJ (1995) Increases in protein kinase C gamma immunoreactivity in the spinal cord dorsal horn of rats with painful mononeuropathy. Neurosci Lett 198: 75–78. doi: 10.1016/0304-3940(95)11975-3
[24]
Tsuda M, Inoue K, Salter MW (2005) Neuropathic pain and spinal microglia: a big problem from molecules in “small” glia. Trends Neurosci 28: 101–107. doi: 10.1016/j.tins.2004.12.002
[25]
Katsura H, Obata K, Mizushima T, Sakurai J, Kobayashi K, et al. (2006) Activation of Src-family kinases in spinal microglia contributes to mechanical hypersensitivity after nerve injury. J Neurosci 26: 8680–8690. doi: 10.1523/jneurosci.1771-06.2006
Zhuang ZY, Gerner P, Woolf CJ, Ji RR (2005) ERK is sequentially activated in neurons, microglia, and astrocytes by spinal nerve ligation and contributes to mechanical allodynia in this neuropathic pain model. Pain 114: 149–159. doi: 10.1016/j.pain.2004.12.022
[28]
Calvo M, Zhu N, Grist J, Ma Z, Loeb JA, et al.. (2011) Following nerve injury neuregulin-1 drives microglial proliferation and neuropathic pain via the MEK/ERK pathway. Glia.
[29]
Svensson CI, Schafers M, Jones TL, Powell H, Sorkin LS (2005) Spinal blockade of TNF blocks spinal nerve ligation-induced increases in spinal P-p38. Neurosci Lett 379: 209–213. doi: 10.1016/j.neulet.2004.12.064
[30]
Sung CS, Wen ZH, Chang WK, Chan KH, Ho ST, et al. (2005) Inhibition of p38 mitogen-activated protein kinase attenuates interleukin-1beta-induced thermal hyperalgesia and inducible nitric oxide synthase expression in the spinal cord. J Neurochem 94: 742–752. doi: 10.1111/j.1471-4159.2005.03226.x
[31]
Abbadie C, Lindia JA, Cumiskey AM, Peterson LB, Mudgett JS, et al. (2003) Impaired neuropathic pain responses in mice lacking the chemokine receptor CCR2. Proc Natl Acad Sci U S A 100: 7947–7952. doi: 10.1073/pnas.1331358100
[32]
Zhuang ZY, Kawasaki Y, Tan PH, Wen YR, Huang J, et al. (2007) Role of the CX3CR1/p38 MAPK pathway in spinal microglia for the development of neuropathic pain following nerve injury-induced cleavage of fractalkine. Brain Behav Immun 21: 642–651. doi: 10.1016/j.bbi.2006.11.003
[33]
Tang Q, Svensson CI, Fitzsimmons B, Webb M, Yaksh TL, et al. (2007) Inhibition of spinal constitutive NOS-2 by 1400W attenuates tissue injury and inflammation-induced hyperalgesia and spinal p38 activation. Eur J Neurosci 25: 2964–2972. doi: 10.1111/j.1460-9568.2007.05576.x
[34]
Clark AK, Malcangio M (2012) Microglial signalling mechanisms: Cathepsin S and Fractalkine. Exp Neurol 234: 283–292. doi: 10.1016/j.expneurol.2011.09.012
[35]
Ji RR, Xu ZZ, Wang X, Lo EH (2009) Matrix metalloprotease regulation of neuropathic pain. Trends Pharmacol Sci 30: 336–340. doi: 10.1016/j.tips.2009.04.002
[36]
Svensson CI, Marsala M, Westerlund A, Calcutt NA, Campana WM, et al. (2003) Activation of p38 mitogen-activated protein kinase in spinal microglia is a critical link in inflammation-induced spinal pain processing. J Neurochem 86: 1534–1544. doi: 10.1046/j.1471-4159.2003.01969.x
[37]
Ji RR, Suter MR (2007) p38 MAPK, microglial signaling, and neuropathic pain. Mol Pain 3: 33. doi: 10.1186/1744-8069-3-33
[38]
Wen YR, Tan PH, Cheng JK, Liu YC, Ji RR (2011) Microglia: a promising target for treating neuropathic and postoperative pain, and morphine tolerance. J Formos Med Assoc 110: 487–494. doi: 10.1016/s0929-6646(11)60074-0
[39]
Wolf G, Gabay E, Tal M, Yirmiya R, Shavit Y (2006) Genetic impairment of interleukin-1 signaling attenuates neuropathic pain, autotomy, and spontaneous ectopic neuronal activity, following nerve injury in mice. Pain 120: 315–324. doi: 10.1016/j.pain.2005.11.011
[40]
Maione S, Siniscalco D, Galderisi U, de Novellis V, Uliano R, et al. (2002) Apoptotic genes expression in the lumbar dorsal horn in a model neuropathic pain in rat. Neuroreport 13: 101–106. doi: 10.1097/00001756-200201210-00024
[41]
Moore KA, Kohno T, Karchewski LA, Scholz J, Baba H, et al. (2002) Partial peripheral nerve injury promotes a selective loss of GABAergic inhibition in the superficial dorsal horn of the spinal cord. J Neurosci 22: 6724–6731.
[42]
Whiteside GT, Munglani R (2001) Cell death in the superficial dorsal horn in a model of neuropathic pain. J Neurosci Res 64: 168–173. doi: 10.1002/jnr.1062
[43]
Campana WM, Myers RR (2003) Exogenous erythropoietin protects against dorsal root ganglion apoptosis and pain following peripheral nerve injury. Eur J Neurosci 18: 1497–1506. doi: 10.1046/j.1460-9568.2003.02875.x
[44]
Sekiguchi M, Kobayashi H, Sekiguchi Y, Konno S, Kikuchi S (2008) Sympathectomy reduces mechanical allodynia, tumor necrosis factor-alpha expression, and dorsal root ganglion apoptosis following nerve root crush injury. Spine (Phila Pa 1976) 33: 1163–1169. doi: 10.1097/brs.0b013e31817144fc
[45]
Schaeffer V, Meyer L, Patte-Mensah C, Eckert A, Mensah-Nyagan AG (2010) Sciatic nerve injury induces apoptosis of dorsal root ganglion satellite glial cells and selectively modifies neurosteroidogenesis in sensory neurons. Glia 58: 169–180. doi: 10.1002/glia.20910
[46]
Mizushima N, Levine B, Cuervo AM, Klionsky DJ (2008) Autophagy fights disease through cellular self-digestion. Nature 451: 1069–1075. doi: 10.1038/nature06639
[47]
Zhang E, Yi MH, Ko Y, Kim HW, Seo JH, et al. (2013) Expression of LC3 and Beclin 1 in the spinal dorsal horn following spinal nerve ligation-induced neuropathic pain. Brain Res 1519: 31–39. doi: 10.1016/j.brainres.2013.04.055
[48]
Zhang W, Sun XF, Bo JH, Zhang J, Liu XJ, et al. (2013) Activation of mTOR in the spinal cord is required for pain hypersensitivity induced by chronic constriction injury in mice. Pharmacol Biochem Behav 111C: 64–70. doi: 10.1016/j.pbb.2013.07.017
