Skip to main content

Advertisement

SpringerLink
Log in

The Domain II S4-S5 Linker in Nav1.9: A Missense Mutation Enhances Activation, Impairs Fast Inactivation, and Produces Human Painful Neuropathy

  • Original Paper
  • Published:
NeuroMolecular Medicine Aims and scope Submit manuscript

Abstract

Painful small fiber neuropathy is a challenging medical condition with no effective treatment. Non-genetic causes can be identified in one half of the subjects. Gain-of-function variants of sodium channels Nav1.7 and Nav1.8 have recently been associated with painful small fiber neuropathy. More recently, mutations of sodium channel Nav1.9 have been linked to human pain disorders, with two gain-of-function mutations found in patients with painful small fiber neuropathy. Here we report a novel Nav1.9 mutation, a glycine 699 substitution by arginine (G699R) in the domain II S4-S5 linker, identified in a patient with painful small fiber neuropathy. In this study, we assayed the mutant channels by voltage-clamp in superior cervical ganglion neurons, which do not produce endogenous Nav1.8 or Nav1.9 currents, and provide a novel platform where Nav1.9 is expressed at relatively high levels. Voltage-clamp analysis showed that the mutation hyperpolarizes (−10.1 mV) channel activation, depolarizes (+6.3 mV) steady-state fast inactivation, slows deactivation, and enhances ramp responses compared with wild-type Nav1.9 channels. Current-clamp analysis showed that the G699R mutant channels render dorsal root ganglion neurons hyperexcitable, via depolarized resting membrane potential, reduced current threshold and increased evoked firing. These observations show that the domain II S4-S5 linker plays an important role in the gating of Nav1.9 and demonstrates that a mutation in this linker is linked to a common pain disorder.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

References

  • Allen, A. S., Berkovic, S. F., Cossette, P., Delanty, N., Dlugos, D., Eichler, E. E., et al. (2013). De novo mutations in epileptic encephalopathies. Nature, 501(7466), 217–221. doi:10.1038/Nature12439.

    Article  CAS  PubMed  Google Scholar 

  • Amaya, F., Wang, H., Costigan, M., Allchorne, A. J., Hatcher, J. P., Egerton, J., et al. (2006). The voltage-gated sodium channel Na(v)1.9 is an effector of peripheral inflammatory pain hypersensitivity. Journal of Neuroscience, 26(50), 12852–12860. doi:10.1523/JNEUROSCI.4015-06.2006.

    Article  CAS  PubMed  Google Scholar 

  • Bendahhou, S., Cummins, T. R., Kula, R. W., Fu, Y. H., & Ptacek, L. J. (2002). Impairment of slow inactivation as a common mechanism for periodic paralysis in DIIS4-S5. Neurology, 58(8), 1266–1272.

    Article  CAS  PubMed  Google Scholar 

  • Cannon, S. C. (2002). An expanding view for the molecular basis of familial periodic paralysis. Neuromuscular Disorders, 12(6), 533–543. doi:10.1016/S0960-8966(02)00007-X.

    Article  PubMed  Google Scholar 

  • Cummins, T. R., Dib-Hajj, S. D., Black, J. A., Akopian, A. N., Wood, J. N., & Waxman, S. G. (1999). A novel persistent tetrodotoxin-resistant sodium current in SNS-null and wild-type small primary sensory neurons. Journal of neuroscience, 19(24).

  • Cummins, T. R., Dib-Hajj, S. D., & Waxman, S. G. (2004). Electrophysiological properties of mutant Nav1.7 sodium channels in a painful inherited neuropathy. Journal of Neuroscience, 24(38), 8232–8236. doi:10.1523/JNEUROSCI.2695-04.2004.

    Article  CAS  PubMed  Google Scholar 

  • Dib-Hajj, S., Black, J. A., Cummins, T. R., & Waxman, S. G. (2002). NaN/Nav1.9: a sodium channel with unique properties. Trends in Neurosciences, 25(5), 253–259.

    Article  CAS  PubMed  Google Scholar 

  • Dib-Hajj, S. D., Choi, J. S., Macala, L. J., Tyrrell, L., Black, J. A., Cummins, T. R., et al. (2009). Transfection of rat or mouse neurons by biolistics or electroporation. Nature Protocols, 4(8), 1118–1126. doi:10.1038/nprot.2009.90.

    Article  CAS  PubMed  Google Scholar 

  • Dib-Hajj, S. D., Tyrrell, L., Black, J. A., & Waxman, S. G. (1998). NaN, a novel voltage-gated Na channel, is expressed preferentially in peripheral sensory neurons and down-regulated after axotomy. Proceedings of the National Academy of Sciences of the United States of America, 95(15), 8963–8968.

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Dib-Hajj, S. D., Tyrrell, L., Cummins, T. R., Black, J. A., Wood, P. M., & Waxman, S. G. (1999a). Two tetrodotoxin-resistant sodium channels in human dorsal root ganglion neurons. FEBS Letters, 462(1–2), 117–120.

    Article  CAS  PubMed  Google Scholar 

  • Dib-Hajj, S. D., Tyrrell, L., Escayg, A., Wood, P. M., Meisler, M. H., & Waxman, S. G. (1999b). Coding sequence, genomic organization, and conserved chromosomal localization of the mouse gene Scn11a encoding the sodium channel NaN. Genomics, 59(3), 309–318. doi:10.1006/geno.1999.5890.

    Article  CAS  PubMed  Google Scholar 

  • Dib-Hajj, S. D., Yang, Y., Black, J. A., & Waxman, S. G. (2013). The Na(V)1.7 sodium channel: from molecule to man. Nature Reviews Neuroscience, 14(1), 49–62. doi:10.1038/nrn3404.

    Article  CAS  PubMed  Google Scholar 

  • Drenth, J. P., & Waxman, S. G. (2007). Mutations in sodium-channel gene SCN9A cause a spectrum of human genetic pain disorders. The Journal of Clinical Investigation, 117(12), 3603–3609. doi:10.1172/JCI33297.

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Estacion, M., & Waxman, S. G. (2013). The response of Na(V)1.3 sodium channels to ramp stimuli: multiple components and mechanisms. Journal of Neurophysiology, 109(2), 306–314. doi:10.1152/jn.00438.2012.

    Article  CAS  PubMed  Google Scholar 

  • Faber, C. G., Hoeijmakers, J. G. J., Ahn, H. S., Cheng, X. Y., Han, C. Y., Choi, J. S., et al. (2012a). Gain of function NaV1.7 mutations in idiopathic small fiber neuropathy. Annals of Neurology, 71(1), 26–39. doi:10.1002/Ana.22485.

    Article  CAS  PubMed  Google Scholar 

  • Faber, C. G., Lauria, G., Merkies, I. S. J., Cheng, X. Y., Han, C. Y., Ahn, H. S., et al. (2012b). Gain-of-function Na(v)1.8 mutations in painful neuropathy. Proceedings of the National Academy of Sciences of the United States of America, 109(47), 19444–19449. doi:10.1073/pnas.1216080109.

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Fjell, J., Hjelmstrom, P., Hormuzdiar, W., Milenkovic, M., Aglieco, F., Tyrrell, L., et al. (2000). Localization of the tetrodotoxin-resistant sodium channel NaN in nociceptors. NeuroReport, 11(1), 199–202. doi:10.1097/00001756-200001170-00039.

    Article  CAS  PubMed  Google Scholar 

  • Han, C., Hoeijmakers, J. G., Ahn, H. S., Zhao, P., Shah, P., Lauria, G., et al. (2012). Nav1.7-related small fiber neuropathy: impaired slow-inactivation and DRG neuron hyperexcitability. Neurology, 78(21), 1635–1643. doi:10.1212/WNL.0b013e3182574f12.

    Article  CAS  PubMed  Google Scholar 

  • Han, C., Rush, A. M., Dib-Hajj, S. D., Li, S., Xu, Z., Wang, Y., et al. (2006). Sporadic onset of erythermalgia: a gain-of-function mutation in Nav1.7. Annals of Neurology, 59(3), 553–558. doi:10.1002/ana.20776.

    Article  CAS  PubMed  Google Scholar 

  • Han, C., Vasylyev, D., Macala, L. J., Gerrits, M. M., Hoeijmakers, J. G., Bekelaar, K. J., et al. (2014). The G1662S NaV1.8 mutation in small fibre neuropathy: impaired inactivation underlying DRG neuron hyperexcitability. Journal of Neurology, Neurosurgery and Psychiatry, 85(5), 499–505. doi:10.1136/jnnp-2013-306095.

    Article  PubMed  Google Scholar 

  • Hockley, J. R., Boundouki, G., Cibert-Goton, V., McGuire, C., Yip, P. K., Chan, C., et al. (2014). Multiple roles for NaV1.9 in the activation of visceral afferents by noxious inflammatory, mechanical, and human disease-derived stimuli. Pain, 155(10), 1962–1975, doi:10.1016/j.pain.2014.06.015.

  • Hoeijmakers, J. G., Faber, C. G., Lauria, G., Merkies, I. S., & Waxman, S. G. (2012a). Small-fibre neuropathies-advances in diagnosis, pathophysiology and management. Nature Reviews Neurology, 8(7), 369–379. doi:10.1038/nrneurol.2012.97.

    Article  CAS  PubMed  Google Scholar 

  • Hoeijmakers, J. G., Han, C., Merkies, I. S., Macala, L. J., Lauria, G., Gerrits, M. M., et al. (2012b) Small nerve fibres, small hands and small feet: a new syndrome of pain, dysautonomia and acromesomelia in a kindred with a novel NaV1.7 mutation. Brain, 135(Pt 2), 345–358, doi:10.1093/brain/awr349

  • Huang, J., Han, C., Estacion, M., Vasylyev, D., Hoeijmakers, J. G., Gerrits, M. M., et al. (2014). Gain-of-function mutations in sodium channel Na(V)1.9 in painful neuropathy. Brain, 137, 1627–1642. doi:10.1093/Brain/Awu079.

    Article  PubMed  Google Scholar 

  • Huang, J., Yang, Y., Zhao, P., Gerrits, M. M., Hoeijmakers, J. G., Bekelaar, K., et al. (2013). Small-fiber neuropathy Nav1.8 mutation shifts activation to hyperpolarized potentials and increases excitability of dorsal root ganglion neurons. Journal of Neuroscience, 33(35), 14087–14097. doi:10.1523/JNEUROSCI.2710-13.2013.

    Article  CAS  PubMed  Google Scholar 

  • Lauria, G., Merkies, I. S., & Faber, C. G. (2012). Small fibre neuropathy. Current Opinion in Neurology, 25(5), 542–549. doi:10.1097/WCO.0b013e32835804c5.

    Article  CAS  PubMed  Google Scholar 

  • Leipold, E., Liebmann, L., Korenke, G. C., Heinrich, T., Giesselmann, S., Baets, J., et al. (2013). A de novo gain-of-function mutation in SCN11A causes loss of pain perception. Nature Genetics, 45(11), 1399–1404. doi:10.1038/ng.2767.

    Article  CAS  PubMed  Google Scholar 

  • Lerche, H., Peter, W., Fleischha, R., Pika-Hartlaub, U., Malina, T., Mitrovic, N., et al. (1997). Role in fast inactivation of the IV/S4-S5 loop of the human muscle Na+ channel probed by cysteine mutagenesis. Journal of Physiology, 505(2), 345–352.

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Lolignier, S., Amsalem, M., Maingret, F., Padilla, F., Gabriac, M., Chapuy, E., et al. (2011). Nav1.9 channel contributes to mechanical and heat pain hypersensitivity induced by subacute and chronic inflammation. PLoS ONE, 6(8), e23083. doi:10.1371/journal.pone.0023083.

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • McPhee, J. C., Ragsdale, D. S., Scheuer, T., & Catterall, W. A. (1998). A critical role for the S4-S5 intracellular loop in domain IV of the sodium channel alpha-subunit in fast inactivation. The Journal of biological chemistry, 273(2), 1121–1129.

    Article  CAS  PubMed  Google Scholar 

  • Niimura, H., Matsunaga, A., Kumagai, K., Ohwaki, K., Ogawa, M., Noguchi, H., et al. (2004). Genetic analysis of Brugada syndrome in western Japan - Two novel mutations. Circulation Journal, 68(8), 740–746. doi:10.1253/Circj.68.740.

    Article  CAS  PubMed  Google Scholar 

  • Ostman, J. A., Nassar, M. A., Wood, J. N., & Baker, M. D. (2008). GTP up-regulated persistent Na+ current and enhanced nociceptor excitability require NaV1.9. Journal of Physiology, 586(4), 1077–1087. doi:10.1113/jphysiol.2007.147942.

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Payandeh, J., El-Din, T. M. G., Scheuer, T., Zheng, N., & Catterall, W. A. (2012). Crystal structure of a voltage-gated sodium channel in two potentially inactivated states. Nature, 486(7401), 135–139. doi:10.1038/Nature11077.

    PubMed Central  CAS  PubMed  Google Scholar 

  • Payandeh, J., Scheuer, T., Zheng, N., & Catterall, W. A. (2011). The crystal structure of a voltage-gated sodium channel. Nature, 475(7356), 353–358. doi:10.1038/Nature10238.

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Persson, A. K., Black, J. A., Gasser, A., Cheng, X. Y., Fischer, T. Z., & Waxman, S. G. (2010). Sodium-calcium exchanger and multiple sodium channel isoforms in intra-epidermal nerve terminals. Molecular Pain, 6, doi:10.1186/1744-8069-6-84

  • Plassart-Schiess, E., Lhuillier, L., George, A. L., Fontaine, B., & Tabti, N. (1998). Functional expression of the Ile693Thr Na+ channel mutation associated with paramyotonia congenita in a human cell line. Journal of Physiology, 507(3), 721–727. doi:10.1111/j.1469-7793.1998.721bs.x.

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Priest, B. T., Murphy, B. A., Lindia, J. A., Diaz, C., Abbadie, C., Ritter, A. M., et al. (2005). Contribution of the tetrodotoxin-resistant voltage-gated sodium channel NaV1.9 to sensory transmission and nociceptive behavior. Proceedings of the National Academy of Sciences of the United States of America, 102(26), 9382–9387. doi:10.1073/pnas.0501549102.

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Qiu, F., Jiang, Y., Zhang, H., Liu, Y., & Mi, W. (2012). Increased expression of tetrodotoxin-resistant sodium channels Nav1.8 and Nav1.9 within dorsal root ganglia in a rat model of bone cancer pain. Neuroscience Letters, 512(2), 61–66. doi:10.1016/j.neulet.2012.01.069.

    Article  CAS  PubMed  Google Scholar 

  • Rugiero, F., Mistry, M., Sage, D., Black, J. A., Waxman, S. G., Crest, M., et al. (2003). Selective expression of a persistent tetrodotoxin-resistant Na+ current and NaV1.9 subunit in myenteric sensory neurons. Journal of Neuroscience, 23(7), 2715–2725.

    CAS  PubMed  Google Scholar 

  • Rush, A. M., Dib-Hajj, S. D., Liu, S., Cummins, T. R., Black, J. A., & Waxman, S. G. (2006). A single sodium channel mutation produces hyper- or hypoexcitability in different types of neurons. Proceedings of the National Academy of Sciences of the United States of America, 103(21), 8245–8250. doi:10.1073/pnas.0602813103.

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Smith, M. R., & Goldin, A. L. (1997). Interaction between the sodium channel inactivation linker and domain III S4-S5. Biophysical Journal, 73(4), 1885–1895.

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Waxman, S. G., & Zamponi, G. W. (2014). Regulating excitability of peripheral afferents: emerging ion channel targets. Nature Neuroscience, 17(2), 153–163. doi:10.1038/nn.3602.

    Article  CAS  PubMed  Google Scholar 

  • Yang, Y., Wang, Y., Li, S., Xu, Z., Li, H., Ma, L., et al. (2004). Mutations in SCN9A, encoding a sodium channel alpha subunit, in patients with primary erythermalgia. Journal of Medical Genetics, 41(3), 171–174. doi:10.1136/jmg.2003.012153.

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Zhang, X., Ren, W. L., DeCaen, P., Yan, C. Y., Tao, X., Tang, L., et al. (2012). Crystal structure of an orthologue of the NaChBac voltage-gated sodium channel. Nature, 486(7401), 130–134. doi:10.1038/Nature11054.

    PubMed Central  CAS  PubMed  Google Scholar 

  • Zhang, X. Y., Wen, J., Yang, W., Wang, C., Gao, L., Zheng, L. H., et al. (2013). Gain-of-function mutations in SCN11A cause familial episodic pain. American Journal of Human Genetics, 93(5), 957–966. doi:10.1016/j.ajhg.2013.09.016.

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Zuberi, S. M., Brunklaus, A., Birch, R., Reavey, E., Duncan, J., & Forbes, G. H. (2011). Genotype-phenotype associations in SCN1A-related epilepsies. Neurology, 76(7), 594–600. doi:10.1212/Wnl.0b013e31820c309b.

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

We thank Lawrence J. Macala, Fadia Dib-Hajj, and Palak Shah for technical assistance. We thank Dr. Mark Estacion and Dr. Jianying Huang for valuable comments. This work was supported in part by Grants from the Rehabilitation Research Service and Medical Research Service, Department of Veterans Affairs (SGW and SDH), and Grant #602273 from the European Union Seventh Framework Programme FP7/2007-2013. The Center for Neuroscience & Regeneration Research is a Collaboration of the Paralyzed Veterans of America with Yale University.

Conflict of interest

The authors report no conflicts of interest.

Author information

Authors and Affiliations

  1. Department of Neurology, Yale University School of Medicine, New Haven, CT, 06510, USA

    Chongyang Han, Yang Yang, Jian Qu, Sulayman D. Dib-Hajj & Stephen G. Waxman

  2. Center for Neuroscience and Regeneration Research, Veterans Affairs Medical Center, Bldg. 34, VA Connecticut Healthcare System, 950 Campbell Avenue, West Haven, CT, 06516, USA

    Chongyang Han, Yang Yang, Jian Qu, Sulayman D. Dib-Hajj & Stephen G. Waxman

  3. Department of Neurology, University Medical Centre Maastricht, Maastricht, The Netherlands

    Bianca T. A. de Greef, Janneke G. J. Hoeijmakers, Ingemar S. J. Merkies & Catharina G. Faber

  4. Department of Clinical Genetics, University Medical Centre Maastricht, Maastricht, The Netherlands

    Monique M. Gerrits

  5. Department of Neurology, Amsterdam Medical Center, Amsterdam, The Netherlands

    Camiel Verhamme

  6. Neuromuscular Diseases Unit and Bioinformatics Unit, IRCCS Foundation, “Carlo Besta”, Milan, Italy

    Giuseppe Lauria

  7. Department of Neurology, Spaarne Hospital, Hoofddorp, The Netherlands

    Ingemar S. J. Merkies

Authors
  1. Chongyang Han
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Yang Yang
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Bianca T. A. de Greef
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Janneke G. J. Hoeijmakers
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Monique M. Gerrits
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Camiel Verhamme
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Jian Qu
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Giuseppe Lauria
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Ingemar S. J. Merkies
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Catharina G. Faber
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Sulayman D. Dib-Hajj
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. Stephen G. Waxman
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Stephen G. Waxman.

Additional information

On behalf of the PROPANE Study Group.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Han, C., Yang, Y., de Greef, B.T.A. et al. The Domain II S4-S5 Linker in Nav1.9: A Missense Mutation Enhances Activation, Impairs Fast Inactivation, and Produces Human Painful Neuropathy. Neuromol Med 17, 158–169 (2015). https://doi.org/10.1007/s12017-015-8347-9

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12017-015-8347-9

Keywords

Search

Navigation