Skip to main content
SpringerLink
Log in

Pharmacological properties of neuronal TTX-resistant sodium channels and the role of a critical serine pore residue

  • Ion Channels, Transporters
  • Published:
Pflügers Archiv Aims and scope Submit manuscript

Abstract

Voltage-gated sodium channels can be characterized by their sensitivity to inhibitors. Nav1.5 is sensitive to block by cadmium and extracellular QX-314, but relatively insensitive to tetrodotoxin and saxitoxin. Nav1.4 is tetrodotoxin- and saxitoxin-sensitive but resistant to cadmium and extracellular QX-314. Nav1.8 and Nav1.9 generate slowly inactivating (ITTXr-Slow) and persistent (ITTXr-Per) currents in sensory neurons that are tetrodotoxin-resistant. Tetrodotoxin sensitivity is largely determined by the identity of a single residue; tyrosine 401 in Nav1.4, cysteine 374 in Nav1.5 and serine 356 and 355 in Nav1.8 and Nav1.9. We asked whether Nav1.8 and Nav1.9 share other pharmacological properties as a result of this serine residue. ITTXr-Slow and ITTXr-Per were saxitoxin-resistant and resistant to internal QX-314. ITTXr-Slow was also resistant to external QX-314 and displayed a approximately fourfold higher sensitivity than ITTXr-Per to cadmium. The impact of the serine residue was investigated by replacing tyrosine 401 in Nav1.4 with serine (Y401S) or cysteine (Y401C). Both mutants were resistant to tetrodotoxin and saxitoxin. Whereas Nav1.4-Y401C displayed an increased sensitivity to cadmium and extracellular QX-314, the serine substitution did not alter the sensitivity of Nav1.4 to cadmium or QX-314. Our data indicates that while the serine residue determines the sensitivity of ITTXr-Slow and ITTXr-Per to tetrodotoxin and saxitoxin, it does not determine their insensitivity to QX-314 or their differential sensitivities to cadmium.

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
Fig. 6

Similar content being viewed by others

Abbreviations

Nav1.4:

Voltage-gated sodium channel α-subunit from skeletal muscle

Nav1.8:

SNS or PN3

Nav1.9:

NaN or SNS2

NGF:

Nerve growth factor

BDNF:

Brain-derived neurotrophic factor

NT4/5:

Neurotrophin 4/5

TrkB:

Tyrosine kinase B

TTX:

tetrodotoxin

STX:

Saxitoxin

References

  1. Akopian AN, Sivilotti L, Wood JN (1996) A tetrodotoxin-resistant voltage-gated sodium channel expressed by sensory neurons. Nature 379:257–62

    Article  CAS  PubMed  Google Scholar 

  2. Akopian AN, Souslova V, England S, Okuse K, Ogata N, Ure J, Smith A, Kerr BJ, McMahon SB, Boyce S, Hill R, Stanfa LC, Dickenson AH, Wood JN (1999) The tetrodotoxin-resistant sodium channel SNS has a specialized function in pain pathways. Nat Neurosci 2:541–8

    Google Scholar 

  3. Backx PH, Yue DT, Lawrence JH, Marban E, Tomaselli GF (1992) Molecular localization of an ion-binding site within the pore of mammalian sodium channels. Science 257:248–51

    CAS  PubMed  Google Scholar 

  4. Baker MD, Wood JN (2001) Involvement of Na+ channels in pain pathways. Trends Pharmacol Sci 22:27–31

    Google Scholar 

  5. Blum R, Kafitz KW, Konnerth A (2002) Neurotrophin-evoked depolarization requires the sodium channel Nav1.9. Nature 419:687–693

    Article  CAS  PubMed  Google Scholar 

  6. Brau ME, Elliott JR (1998) Local anaesthetic effects on tetrodotoxin-resistant Na+ currents in rat dorsal root ganglion neurones. Eur J Anaesthesiol 15:80–88

    Article  CAS  PubMed  Google Scholar 

  7. Carbonneau E, Vijayaragavan K, Chahine M (2002) A tryptophan residue (W736) in the amino-terminus of the P-segment of domain II is involved in pore formation in Nav1.4 voltage-gated sodium channels. Pflugers Arch 445:18–24

    Article  CAS  PubMed  Google Scholar 

  8. Cummins TR, Black JA, Dib-Hajj SD, Waxman SG (2000) Glial-derived neurotrophic factor upregulates expression of functional SNS and NaN sodium channels and their currents in axotomized dorsal root ganglion neurons. J Neurosci 20:8754–8761

    CAS  PubMed  Google Scholar 

  9. Cummins TR, Dib-Hajj SD, Black JA, Akopian AN, Wood JN, Waxman SG (1999) A novel persistent tetrodotoxin-resistant sodium current in SNS-null and wild-type small primary sensory neurons. J Neurosci 19:RC43, 1–6

    CAS  PubMed  Google Scholar 

  10. Cummins TR, Howe JR, Waxman SG (1998) Slow closed-state inactivation underlies tetrodotoxin-sensitive ramp currents in HEK293 cells expressing hNE sodium channels and in dorsal root ganglion neurons. J Neurosci 18:9607–9619

    CAS  PubMed  Google Scholar 

  11. Cummins TR, Waxman SG (1997) Downregulation of tetrodotoxin-resistant sodium currents and upregulation of a rapidly repriming tetrodotoxin-sensitive sodium current in small spinal sensory neurons after nerve injury. J Neurosci 17:3503–3514

    CAS  PubMed  Google Scholar 

  12. Dib-Hajj S, Black JA, Cummins TR, Waxman SG (2002) NaN/Nav1.9: a sodium channel with unique properties. Trends Neurosci 25:253–259

    Article  CAS  PubMed  Google Scholar 

  13. Dib-Hajj SD, Tyrrell L, Black JA, Waxman SG (1998) NaN, a novel voltage-gated Na channel, is expressed preferentially in peripheral sensory neurons and down-regulated after axotomy. Proc Natl Acad Sci USA 95:8963–8968

    Article  CAS  PubMed  Google Scholar 

  14. Dib-Hajj SD, Tyrrell L, Cummins TR, Black JA, Wood PM, Waxman SG (1999) Two tetrodotoxin-resistant sodium channels in human dorsal root ganglion neurons. FEBS Lett 462:117–120

    Article  CAS  PubMed  Google Scholar 

  15. Favre I, Moczydlowski E, Schild L (1995) Specificity for block by saxitoxin and divalent cations at a residue which determines sensitivity of sodium channel subtypes to guanidinium toxins. J Gen Physiol 106:203–229

    Article  CAS  PubMed  Google Scholar 

  16. Frelin C, Cognard C, Vigne P, Lazdunski M (1986) Tetrodotoxin-sensitive and tetrodotoxin-resistant Na+ channels differ in their sensitivity to Cd2+ and Zn2+. Eur J Pharmacol 122:245–250

    Article  CAS  PubMed  Google Scholar 

  17. Heinemann SH, Terlau H, Imoto K (1992) Molecular basis for pharmacological differences between brain and cardiac sodium channels. Pflugers Arch 422:90–92

    Article  CAS  PubMed  Google Scholar 

  18. Herzog RI, Cummins TR, Waxman SG (2001) Persistent TTX-resistant Na+ current affects resting potential and response to depolarization in simulated spinal sensory neurons. J Neurophysiol 86:1351–1364

    CAS  PubMed  Google Scholar 

  19. Liu H, Atkins J, Kass RS (2003) Common molecular determinants of flecainide and lidocaine block of heart Na+ channels: evidence from experiments with neutral and quaternary flecainide analogues. J Gen Physiol 121:199–214

    Article  CAS  PubMed  Google Scholar 

  20. Murphy BA, Abbadie C, Kaczorowski GJ, Lindia JA, Priest BT, Ritter AM, Martin WJ (2004) Contribution of the tetrodotoxin-resistant sodium channel Nav1.9 to sensory transmission and nociceptive behavior in mouse. Soc Neurosci Abstr 62.1, 2004

  21. Qu Y, Rogers J, Tanada T, Scheuer T, Catterall WA (1995) Molecular determinants of drug access to the receptor site for antiarrhythmic drugs in the cardiac Na+ channel. Proc Natl Acad Sci USA 92:11839–11843

    CAS  PubMed  Google Scholar 

  22. Ragsdale DS, McPhee JC, Scheuer T, Catterall WA (1994) Molecular determinants of state-dependent block of Na+ channels by local anesthetics. Science 265:1724–1728

    CAS  PubMed  Google Scholar 

  23. Renganathan M, Cummins TR, Waxman SG (2001) Contribution of Nav1.8 sodium channels to action potential electrogenesis in DRG neurons. J Neurophysiol 86:629–640

    CAS  PubMed  Google Scholar 

  24. Roy ML, Narahashi T (1992) Differential properties of tetrodotoxin-sensitive and tetrodotoxin-resistant sodium channels in rat dorsal root ganglion neurons. J Neurosci 12:2104–2111

    CAS  PubMed  Google Scholar 

  25. Rush AM, Elliott JR (1997) Phenytoin and carbamazepine: differential inhibition of sodium currents in small cells from adult rat dorsal root ganglia. Neurosci Lett 226:95–98

    Article  CAS  PubMed  Google Scholar 

  26. Satin J, Kyle JW, Chen M, Bell P, Cribbs LL, Fozzard HA, Rogart RB (1992) A mutant of TTX-resistant cardiac sodium channels with TTX-sensitive properties. Science 256:1202–1205

    CAS  PubMed  Google Scholar 

  27. Sivilotti L, Okuse K, Akopian AN, Moss S, Wood JN (1997) A single serine residue confers tetrodotoxin insensitivity on the rat sensory-neuron-specific sodium channel SNS. FEBS Lett 409:49–52

    Article  CAS  PubMed  Google Scholar 

  28. Sleeper AA, Cummins TR, Dib-Hajj SD, Hormuzdiar W, Tyrrell L, Waxman SG, Black JA (2000) Changes in expression of two tetrodotoxin-resistant sodium channels and their currents in dorsal root ganglion neurons after sciatic nerve injury but not rhizotomy. J Neurosci 20:7279–7289

    CAS  PubMed  Google Scholar 

  29. Sunami A, Glaaser IW, Fozzard HA (2000) A critical residue for isoform difference in tetrodotoxin affinity is a molecular determinant of the external access path for local anesthetics in the cardiac sodium channel. Proc Natl Acad Sci USA 97:2326–2331

    Article  CAS  PubMed  Google Scholar 

  30. Tate S, Benn S, Hick C, Trezise D, John V, Mannion RJ, Costigan M, Plumpton C, Grose D, Gladwell Z, Kendall G, Dale K, Bountra C, Woolf CJ (1998) Two sodium channels contribute to the TTX-R sodium current in primary sensory neurons. Nat Neurosci 1:653–655

    Article  CAS  PubMed  Google Scholar 

  31. Ukomadu C, Zhou J, Sigworth FJ, Agnew WS (1992) muI Na+ channels expressed transiently in human embryonic kidney cells: biochemical and biophysical properties. Neuron 8:663–676

    Article  CAS  PubMed  Google Scholar 

  32. Wang SY, Nau C, Wang GK (2000) Residues in Na(+) channel D3-S6 segment modulate both batrachotoxin and local anesthetic affinities. Biophys J 79:1379–1387

    CAS  PubMed  Google Scholar 

  33. Waxman SG, Dib-Hajj S, Cummins TR, Black JA (1999) Sodium channels and pain. Proc Natl Acad Sci USA 96:7635–7639

    Article  CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Klinik für Anästhesiologie, Friedrich-Alexander-Universität Erlangen-Nuremberg, Krankenhausstr. 12, 91054, Erlangen, Germany

    Andreas Leffler

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

    Raimund I. Herzog, Sulayman D. Dib-Hajj & Stephen G. Waxman

  3. Department of Pharmacology and Toxicology, Stark Neurosciences Research Center, Indiana University School of Medicine, R2-402 950 W Walnut Street, Indianapolis, IN, 46202, USA

    Theodore R. Cummins

Authors
  1. Andreas Leffler
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Raimund I. Herzog
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Sulayman D. Dib-Hajj
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Stephen G. Waxman
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Theodore R. Cummins
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Theodore R. Cummins.

Additional information

This work was supported in part by grants from the Medical Research Service and the Rehabilitation Research Service, Department of Veterans Affairs, and by grants from The Eastern Paralyzed Veterans Association and the Paralyzed Veterans of America.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Leffler, A., Herzog, R.I., Dib-Hajj, S.D. et al. Pharmacological properties of neuronal TTX-resistant sodium channels and the role of a critical serine pore residue. Pflugers Arch - Eur J Physiol 451, 454–463 (2005). https://doi.org/10.1007/s00424-005-1463-x

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00424-005-1463-x

Keywords

Search

Navigation