Molecular Medicine

, Volume 21, Issue 1, pp 544–552 | Cite as

Virus-Mediated Knockdown of Nav1.3 in Dorsal Root Ganglia of STZ-Induced Diabetic Rats Alleviates Tactile Allodynia

  • Andrew M. Tan
  • Omar A. Samad
  • Sulayman D. Dib-Hajj
  • Stephen G. Waxman
Research Article


Diabetic neuropathic pain affects a substantial number of people and represents a major public health problem. Available clinical treatments for diabetic neuropathic pain remain only partially effective and many of these treatments carry the burden of side effects or the risk of dependence. The misexpression of sodium channels within nociceptive neurons contributes to abnormal electrical activity associated with neuropathic pain. Voltage-gated sodium channel Nav1.3 produces tetrodotoxin-sensitive sodium currents with rapid repriming kinetics and has been shown to contribute to neuronal hyperexcitability and ectopic firing in injured neurons. Suppression of Nav1.3 activity can attenuate neuropathic pain induced by peripheral nerve injury. Previous studies have shown that expression of Nav1.3 is upregulated in dorsal root ganglion (DRG) neurons of diabetic rats that exhibit neuropathic pain. Here, we hypothesized that viral-mediated knockdown of Nav1.3 in painful diabetic neuropathy would reduce neuropathic pain. We used a validated recombinant adeno-associated virus (AAV)-shRNA-Nav1.3 vector to knockdown expression of Nav1.3, via a clinically applicable intrathecal injection method. Three weeks following vector administration, we observed a significant rate of transduction in DRGs of diabetic rats that concomitantly reduced neuronal excitability of dorsal horn neurons and reduced behavioral evidence of tactile allodynia. Taken together, these findings offer a novel gene therapy approach for addressing chronic diabetic neuropathic pain.



This work was supported by grants from the Nancy Taylor Foundation and the Department of Veterans Affairs (VA) Medical Research Service and Rehabilitation Research Service. AM Tan and OA Samad are funded by the Nancy Taylor Foundation. AM Tan is funded by grants from the PVA Research Foundation and the US Department of Veterans Affairs (1 IK2 RX001123-01A2). The Center for Neuroscience and Regeneration Research is a Collaboration of the Paralyzed Veterans of America with Yale University. We thank Pamela Zwinger, Shujun Liu, and Peng Zhao for their excellent technical assistance.


  1. 1.
    Zilliox L, Russell JW. (2011) Treatment of diabetic sensory polyneuropathy. Curr. Treat. Options Neurol. 13:143–59.CrossRefGoogle Scholar
  2. 2.
    Dib-Hajj SD, Black JA, Waxman SG. (2009) Voltage-gated sodium channels: therapeutic targets for pain. Pain Med. 10:1260–9.CrossRefGoogle Scholar
  3. 3.
    Dib-Hajj SD, Cummins TR, Black JA, Waxman SG. (2010) Sodium channels in normal and pathological pain. Annu. Rev. Neurosci. 33:325–47.CrossRefGoogle Scholar
  4. 4.
    Black JA, et al. (1999) Upregulation of a silent sodium channel after peripheral, but not central, nerve injury in DRG neurons. J. Neurophysiol. 82:2776–85.CrossRefGoogle Scholar
  5. 5.
    Samad OA, et al. (2013) Virus-mediated shRNA knockdown of Na(v)1.3 in rat dorsal root ganglion attenuates nerve injury-induced neuropathic pain. Mol. Ther. 21:49–56.CrossRefGoogle Scholar
  6. 6.
    Waxman SG, Kocsis JD, Black JA. (1994) Type III sodium channel mRNA is expressed in embryonic but not adult spinal sensory neurons, and is reexpressed following axotomy. J. Neurophysiol. 72:466–70.CrossRefGoogle Scholar
  7. 7.
    Cummins TR, et al. (2001) Nav1.3 sodium channels: rapid repriming and slow closed-state inactivation display quantitative differences after expression in a mammalian cell line and in spinal sensory neurons. J. Neurosci. 21:5952–61.CrossRefGoogle Scholar
  8. 8.
    Waxman SG, Hains BC. (2006) Fire and phantoms after spinal cord injury: Na+ channels and central pain. Trends Neurosci. 29:207–15.CrossRefGoogle Scholar
  9. 9.
    Lindia JA, Kohler MG, Martin WJ, Abbadie C. (2005) Relationship between sodium channel NaV1.3 expression and neuropathic pain behavior in rats. Pain. 117:145–53.CrossRefGoogle Scholar
  10. 10.
    Nassar MA, et al. (2006) Nerve injury induces robust allodynia and ectopic discharges in Nav1.3 null mutant mice. Mol. Pain. 2:33.CrossRefGoogle Scholar
  11. 11.
    Yin R, et al. (2015) Voltage-gated sodium channel function and expression in injured and uninjured rat dorsal root ganglia neurons. Int. J. Neurosci. 2015, Apr 7 [Epub ahead of print].Google Scholar
  12. 12.
    Craner MJ, Klein JP, Renganathan M, Black JA, Waxman SG. (2002) Changes of sodium channel expression in experimental painful diabetic neuropathy. Ann. Neurol. 52:786–92.CrossRefGoogle Scholar
  13. 13.
    Shah BS, et al. (2001) Beta3, a novel auxiliary subunit for the voltage gated sodium channel is upregulated in sensory neurones following streptozocin induced diabetic neuropathy in rat. Neurosci. Lett. 309:1–4.CrossRefGoogle Scholar
  14. 14.
    Szolcsanyi J, Pinter E. (2013) Transient receptor potential vanilloid 1 as a therapeutic target in analgesia. Expert Opin. Ther. Targets. 17:641–57.CrossRefGoogle Scholar
  15. 15.
    Glorioso JC, Fink DJ. (2009) Gene therapy for pain: introduction to the special issue. Gene Ther. 16:453–4.CrossRefGoogle Scholar
  16. 16.
    Tan AM, et al. (2012) Maladaptive dendritic spine remodeling contributes to diabetic neuropathic pain. J. Neurosci. 32:6795–807.CrossRefGoogle Scholar
  17. 17.
    Morrow TJ. (2004) Animal models of painful diabetic neuropathy: the STZ rat model. Curr. Protoc. Neurosci. Chapter 9:Unit 9.18.Google Scholar
  18. 18.
    Fischer TZ, Tan AM, Waxman SG. (2009) Thalamic neuron hyperexcitability and enlarged receptive fields in the STZ model of diabetic pain. Brain Res. 1268:154–61.CrossRefGoogle Scholar
  19. 19.
    Persson AK, Gasser A, Black JA, Waxman SG. (2011) Na1.7 accumulates and co-localizes with phosphorylated ERK1/2 within transected axons in early experimental neuromas. Exp. Neurol. 230:273–9.CrossRefGoogle Scholar
  20. 20.
    Chaplan SR, Bach FW, Pogrel JW, Chung JM, Yaksh TL. (1994) Quantitative assessment of tactile allodynia in the rat paw. J. Neurosci. Methods. 53:55–63.CrossRefGoogle Scholar
  21. 21.
    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.CrossRefGoogle Scholar
  22. 22.
    Chang YW, Tan A, Saab C, Waxman S. (2010) Unilateral focal burn injury is followed by long-lasting bilateral allodynia and neuronal hyperexcitability in spinal cord dorsal horn. J. Pain. 11:119–30.CrossRefGoogle Scholar
  23. 23.
    Baumgartner U, Magerl W, Klein T, Hopf HC, Treede RD. (2002) Neurogenic hyperalgesia versus painful hypoalgesia: two distinct mechanisms of neuropathic pain. Pain. 96:141–51.CrossRefGoogle Scholar
  24. 24.
    Pitcher GM, Henry JL. (2004) Nociceptive response to innocuous mechanical stimulation is mediated via myelinated afferents and NK-1 receptor activation in a rat model of neuropathic pain. Exp. Neurol. 186:173–97.CrossRefGoogle Scholar
  25. 25.
    Parikh P, et al. (2011) Regeneration of axons in injured spinal cord by activation of bone morphogenetic protein/Smad1 signaling pathway in adult neurons. Proc. Natl. Acad. Sci U. S. A. 108:E99–107.CrossRefGoogle Scholar
  26. 26.
    Xu Q, et al. (2012) In vivo gene knockdown in rat dorsal root ganglia mediated by self-complementary adeno-associated virus serotype 5 following intrathecal delivery. PloS One 7:e32581.CrossRefGoogle Scholar
  27. 27.
    Tan AM, Chang YW, Zhao P, Hains BC, Waxman SG. (2011) Rac1-regulated dendritic spine remodeling contributes to neuropathic pain after peripheral nerve injury. Exp. Neurol. 232:222–33.CrossRefGoogle Scholar
  28. 28.
    Gjerstad J, Tjolsen A, Hole K. (2001) Induction of long-term potentiation of single wide dynamic range neurones in the dorsal horn is inhibited by descending pathways. Pain. 91:263–8.CrossRefGoogle Scholar
  29. 29.
    Setacci C, de Donato G, Setacci F, Chisci E. (2009) Diabetic patients: epidemiology and global impact. J. Cardiovasc. Surg. 50:263–73.Google Scholar
  30. 30.
    Cummins TR, Waxman SG. (1997) Downregulation of tetrodotoxin-resistant sodium currents and upregulation of a rapidly repriming tetrodotoxinsensitive sodium current in small spinal sensory neurons after nerve injury. J. Neurosci. 17:3503–14.CrossRefGoogle Scholar
  31. 31.
    Pertovaara A, Wei H, Kalmari J, Ruotsalainen M. (2001) Pain behavior and response properties of spinal dorsal horn neurons following experimental diabetic neuropathy in the rat: modulation by nitecapone, a COMT inhibitor with antioxidant properties. Exp. Neurol. 167:425–34.CrossRefGoogle Scholar
  32. 32.
    Raz I, Hasdai D, Seltzer Z, Melmed RN. (1988) Effect of hyperglycemia on pain perception and on efficacy of morphine analgesia in rats. Diabetes. 37:1253–9.CrossRefGoogle Scholar
  33. 33.
    Courteix C, Eschalier A, Lavarenne J. (1993) Streptozocin-induced diabetic rats: behavioural evidence for a model of chronic pain. Pain. 53:81–8.CrossRefGoogle Scholar
  34. 34.
    Benbow SJ, Chan AW, Bowsher D, MacFarlane IA, Williams G. (1994) A prospective study of painful symptoms, small-fibre function and peripheral vascular disease in chronic painful diabetic neuropathy. Diabet. Med. 11:17–21.CrossRefGoogle Scholar
  35. 35.
    Boucek P. (2006) Advanced diabetic neuropathy: a point of no return? Rev. Diabet. Stud. 3:143–50.CrossRefGoogle Scholar
  36. 36.
    Jain R. (2008) Pain and the brain: diabetic neuropathic pain. J. Clin. Psychiatry. 69:e22.CrossRefGoogle Scholar
  37. 37.
    Ahlgren SC, Levine JD. (1994) Protein kinase C inhibitors decrease hyperalgesia and C-fiber hyperexcitability in the streptozotocin-diabetic rat. J. Neurophysiol. 72:684–92.CrossRefGoogle Scholar
  38. 38.
    Chen SR, Pan HL. (2002) Hypersensitivity of spinothalamic tract neurons associated with diabetic neuropathic pain in rats. J. Neurophysiol. 87:2726–33.CrossRefGoogle Scholar
  39. 39.
    Fischer TZ, Waxman SG. (2010) Neuropathic pain in diabetes—evidence for a central mechanism. Nat. Rev. Neurol. 6:462–6.CrossRefGoogle Scholar
  40. 40.
    Hains BC, Saab CY, Waxman SG. (2005) Changes in electrophysiological properties and sodium channel Nav1.3 expression in thalamic neurons after spinal cord injury. Brain. 128:2359–71.CrossRefGoogle Scholar
  41. 41.
    Zhao P, Waxman SG, Hains BC. (2006) Sodium channel expression in the ventral posterolateral nucleus of the thalamus after peripheral nerve injury. Mol. Pain. 2:27.CrossRefGoogle Scholar

Copyright information

© The Author(s) 2015

Open Access This article is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License, which permits any non-commercial use, sharing, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, and provide a link to the Creative Commons license. You do not have permission under this license to share adapted material derived from this article or parts of it.

The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder.

To view a copy of this license, visit (

Authors and Affiliations

  • Andrew M. Tan
    • 1
    • 2
  • Omar A. Samad
    • 1
    • 2
  • Sulayman D. Dib-Hajj
    • 1
    • 2
  • Stephen G. Waxman
    • 1
    • 2
  1. 1.Department of NeurologyYale University School of MedicineNew HavenUSA
  2. 2.The Center for Neuroscience and Regeneration Research, Department of Neurology, Veterans Affairs Connecticut Healthcare SystemYale UniversityWest HavenUSA

Personalised recommendations