Abstract
Veratridine is a lipid-soluble neurotoxin derived from plants in the family Liliaceae. It has been broadly investigated for its action as a sodium channel agonist. However, the effects of veratridine on subtypes of sodium channels, especially Nav1.7, remain to be studied. Here, we investigated the effects of veratridine on human Nav1.7 ectopically expressed in HEK293A cells and recorded Nav1.7 currents from the cells using whole-cell patch clamp technique. We found that veratridine exerted a dose-dependent inhibitory effect on the peak current of Nav1.7, with the half-maximal inhibition concentration (IC50) of 18.39 µM. Meanwhile, veratridine also elicited tail current (linearly) and sustained current [half-maximal concentration (EC50): 9.53 µM], also in a dose-dependent manner. Veratridine (75 µM) shifted the half-maximal activation voltage of the Nav1.7 activation curve in the hyperpolarized direction, from −21.64 ± 0.75 mV to −28.14 ± 0.66 mV, and shifted the half-inactivation voltage of the steady-state inactivation curve from −59.39 ± 0.39 mV to −73.78 ± 0.5 mV. An increased frequency of stimulation decreased the peak and tail currents of Nav1.7 for each pulse along with pulse number, and increased the accumulated tail current at the end of train stimulation. These findings reveal the different modulatory effects of veratridine on the Nav1.7 peak current and tail current.
Similar content being viewed by others
Log in or create a free account to read this content
Gain free access to this article, as well as selected content from this journal and more on nature.com
or
References
Wang SY, Wang GK. Voltage-gated sodium channels as primary targets of diverse lipid-soluble neurotoxins. Cell Signal. 2003;15:151–9.
Nemoto T, Yanagita T, Maruta T, Sugita C, Satoh S, Kanai T, et al. Endothelin-1-induced down-regulation of NaV1.7 expression in adrenal chromaffin cells: attenuation of catecholamine secretion and tau dephosphorylation. FEBS Lett. 2013;587:898–905.
Bicknell RJ, Schofield JG. Inhibition by somatostatin of bovine growth hormone secretion following sodium channel activation. J Physiol. 1981;316:85–96.
Hill AJ, Jones NA, Smith I, Hill CL, Williams CM, Stephens GJ, et al. Voltage-gated sodium (NaV) channel blockade by plant cannabinoids does not confer anticonvulsant effects per se. Neurosci Lett. 2014;566:269–74.
Felix JP, Williams BS, Priest BT, Brochu RM, Dick IE, Warren VA, et al. Functional assay of voltage-gated sodium channels using membrane potential-sensitive dyes. Assay Drug Dev Technol. 2004;2:260–8.
Telinius N, Majgaard J, Kim S, Katballe N, Pahle E, Nielsen J, et al. Voltage-gated sodium channels contribute to action potentials and spontaneous contractility in isolated human lymphatic vessels. J Physiol. 2015;593:3109–22.
Saleh S, Yeung SY, Prestwich S, Pucovsky V, Greenwood I. Electrophysiological and molecular identification of voltage-gated sodium channels in murine vascular myocytes. J Physiol. 2005;568:155–69.
Fort A, Cordaillat M, Thollon C, Salazar G, Mechaly I, Villeneuve N, et al. New insights in the contribution of voltage-gated Na(v) channels to rat aorta contraction. PLoS One. 2009;4:e7360.
Auerbach DS, Grzda KR, Furspan PB, Sato PY, Mironov S, Jalife J. Structural heterogeneity promotes triggered activity, reflection and arrhythmogenesis in cardiomyocyte monolayers. J Physiol. 2011;589:2363–81.
Pinto FM, Ravina CG, Fernandez-Sanchez M, Gallardo-Castro M, Cejudo-Roman A, Candenas L. Molecular and functional characterization of voltage-gated sodium channels in human sperm. Reprod Biol Endocrinol. 2009;7:71.
Tsukamoto T, Chiba Y, Nakazaki A, Ishikawa Y, Nakane Y, Cho Y, et al. Inhibition of veratridine-induced delayed inactivation of the voltage-sensitive sodium channel by synthetic analogs of crambescin B. Bioorg Med Chem Lett. 2017;27:1247–51.
Sutro JB. Kinetics of veratridine action on Na channels of skeletal muscle. J Gen Physiol. 1986;87:1–24.
Catterall WA, Cestele S, Yarov-Yarovoy V, Yu FH, Konoki K, Scheuer T. Voltage-gated ion channels and gating modifier toxins. Toxicon. 2007;49:124–41.
Janiszewski L. The action of toxins on the voltage-gated sodium channel. Pol J Pharmacol Pharm. 1990;42:581–8.
Kanai T, Nemoto T, Yanagita T, Maruta T, Satoh S, Yoshikawa N, et al. Nav1.7 sodium channel-induced Ca2+ influx decreases tau phosphorylation via glycogen synthase kinase-3beta in adrenal chromaffin cells. Neurochem Int. 2009;54:497–505.
Richelson E. Lithium ion entry through the sodium channel of cultured mouse neuroblastoma cells: a biochemical study. Science. 1977;196:1001–2.
Armstrong CM, Bezanilla F. Inactivation of the sodium channel. II. Gating current experiments. J Gen Physiol. 1977;70:567–90.
Khodorov BI. Inactivation of the sodium gating current. Neuroscience. 1979;4:865–76.
Kruger LC, Isom LL. Voltage-gated Na+ channels: not just for conduction. Cold Spring Harb Perspect Biol. 2016;8:a029264
Alicata DA, Rayner MD, Starkus JG. Sodium channel activation mechanisms. Insights from deuterium oxide substitution. Biophys J. 1990;57:745–58.
Kuo CC, Bean BP. Na+ channels must deactivate to recover from inactivation. Neuron. 1994;12:819–29.
Black JA, Frezel N, Dib-Hajj SD, Waxman SG. Expression of Nav1.7 in DRG neurons extends from peripheral terminals in the skin to central preterminal branches and terminals in the dorsal horn. Mol Pain. 2012;8:82.
Catterall WA. Structure and function of voltage-gated ion channels. Annu Rev Biochem. 1995;64:493–531.
Wood JN, Boorman JP, Okuse K, Baker MD. Voltage-gated sodium channels and pain pathways. J Neurobiol. 2004;61:55–71.
Dib-Hajj SD, Yang Y, Black JA, Waxman SG. The Na(V)1.7 sodium channel: from molecule to man. Nat Rev Neurosci. 2013;14:49–62.
Zhao F, Li X, Jin L, Zhang F, Inoue M, Yu B, et al. Development of a rapid throughput assay for identification of hNav1.7 antagonist using unique efficacious sodium channel agonist, antillatoxin. Mar Drugs. 2016; 14(2): md14020036.
Farrag KJ, Bhattacharjee A, Docherty RJ. A comparison of the effects of veratridine on tetrodotoxin-sensitive and tetrodotoxin-resistant sodium channels in isolated rat dorsal root ganglion neurons. Pflugers Arch. 2008;455:929–38.
Mohammed ZA, Doran C, Grundy D, Nassar MA. Veratridine produces distinct calcium response profiles in mouse dorsal root ganglia neurons. Sci Rep. 2017;7:45221.
Leibowitz MD, Sutro JB, Hille B. Voltage-dependent gating of veratridine-modified Na channels. J Gen Physiol. 1986;87:25–46.
Zhu HL, Wassall RD, Takai M, Morinaga H, Nomura M, Cunnane TC, et al. Actions of veratridine on tetrodotoxin-sensitive voltage-gated Na currents, Na1.6, in murine vas deferens myocytes. Br J Pharmacol. 2009;157:1483–93.
Cox JJ, Reimann F, Nicholas AK, Thornton G, Roberts E, Springell K, et al. An SCN9A channelopathy causes congenital inability to experience pain. Nature. 2006;444:894–8.
Chai Z, Wang C, Huang R, Wang Y, Zhang X, Wu Q, et al. CaV2.2 gates calcium-independent but voltage-dependent secretion in mammalian sensory. Neurons Neuron. 2017;96:1317–26 e4.
Zhang B, Zhang XY, Luo PF, Huang W, Zhu FP, Liu T, et al. Action potential-triggered somatic exocytosis in mesencephalic trigeminal nucleus neurons in rat brain slices. J Physiol. 2012;590:753–62.
Talvenheimo JA, Tamkun MM, Catterall WA. Reconstitution of neurotoxin-stimulated sodium transport by the voltage-sensitive sodium channel purified from rat brain. J Biol Chem. 1982;257:11868–71.
Tamkun MM, Talvenheimo JA, Catterall WA. The sodium channel from rat brain. Reconstitution of neurotoxin-activated ion flux and scorpion toxin binding from purified components. J Biol Chem. 1984;259:1676–88.
Bouron A, Reuter H. A role of intracellular Na+ in the regulation of synaptic transmission and turnover of the vesicular pool in cultured hippocampal cells. Neuron. 1996;17:969–78.
Catterall WA. Cellular and molecular biology of voltage-gated sodium channels. Physiol Rev. 1992;72:S15–48.
Ulbricht W, Flacke W. After-potentials and large depolarizations of single nodes of Ranvier treated with veratridine. J Gen Physiol. 1965;48:1035–46.
Ulbricht W. The effect of veratridine on excitable membranes of nerve and muscle. Ergeb Physiol. 1969;61:18–71.
Sumiya Y, Torigoe K, Gerevich Z, Kofalvi A, Vizi ES. Excessive release of [3H] noradrenaline by veratridine and ischemia in spinal cord. Neurochem Int. 2001;39:59–63.
Bradley E, Webb TI, Hollywood MA, Sergeant GP, McHale NG, Thornbury KD. The cardiac sodium current Na(v)1.5 is functionally expressed in rabbit bronchial smooth muscle cells. Am J Physiol Cell Physiol. 2013;305:C427–35.
Abdel-Aziz H, Windeck T, Ploch M, Verspohl EJ. Mode of action of gingerols and shogaols on 5-HT3 receptors: binding studies, cation uptake by the receptor channel and contraction of isolated guinea-pig ileum. Eur J Pharmacol. 2006;530:136–43.
Power KE, Carlin KP, Fedirchuk B. Modulation of voltage-gated sodium channels hyperpolarizes the voltage threshold for activation in spinal motoneurones. Exp Brain Res. 2012;217:311–22.
Barnes S, Hille B. Veratridine modifies open sodium channels. J Gen Physiol. 1988;91:421–43.
Ho C, O’Leary ME. Single-cell analysis of sodium channel expression in dorsal root ganglion neurons. Mol Cell Neurosci. 2011;46:159–66.
Liu XP, Wooltorton JR, Gaboyard-Niay S, Yang FC, Lysakowski A, Eatock RA. Sodium channel diversity in the vestibular ganglion: NaV1.5, NaV1.8, and tetrodotoxin-sensitive currents. J Neurophysiol. 2016;115:2536–55.
Rando TA. Rapid and slow gating of veratridine-modified sodium channels in frog myelinated nerve. J Gen Physiol. 1989;93:43–65.
Wang GK, Quan C, Seaver M, Wang SY. Modification of wild-type and batrachotoxin-resistant muscle mu1 Na+ channels by veratridine. Pflugers Arch. 2000;439:705–13.
Wang GK, Wang SY. Veratridine block of rat skeletal muscle Nav1.4 sodium channels in the inner vestibule. J Physiol. 2003;548:667–75.
Chevrier P, Vijayaragavan K, Chahine M. Differential modulation of Nav1.7 and Nav1.8 peripheral nerve sodium channels by the local anesthetic lidocaine. Br J Pharmacol. 2004;142:576–84.
Joca HC, Vieira DC, Vasconcelos AP, Araujo DA, Cruz JS. Carvacrol modulates voltage-gated sodium channels kinetics in dorsal root ganglia. Eur J Pharmacol. 2015;756:22–9.
Pal K, Gangopadhyay G. Probing kinetic drug binding mechanism in voltage-gated sodium ion channel: open state versus inactive state blockers. Channels (Austin). 2015;9:307–16.
Acknowledgements
This study was sponsored by the National Natural Science Foundation of China (31600950). The hNav1.7 plasmid including the α subunit and the β1/β2 subunits was a generous gift from Dr. Christopher Geoffrey Woods (The Clinical Medical School, University of Cambridge, UK). We appreciate Dr. James Cox’s help and guidance with the shipping of this plasmid. We thank Younus Muhammad for providing language help.
Author contributions
X-yZ performed the experiments and wrote the manuscript; PZ prepared the cell line and drug reagents; R-yB contributed to the plasmid preparation; Y-hG designed the research and revised the paper.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Competing interests
The authors declare no competing interest.
Electronic supplementary material
Rights and permissions
About this article
Cite this article
Zhang, Xy., Bi, Ry., Zhang, P. et al. Veratridine modifies the gating of human voltage-gated sodium channel Nav1.7. Acta Pharmacol Sin 39, 1716–1724 (2018). https://doi.org/10.1038/s41401-018-0065-z
Received:
Accepted:
Published:
Version of record:
Issue date:
DOI: https://doi.org/10.1038/s41401-018-0065-z
Keywords
This article is cited by
-
Poneratoxin as a key tool for investigating the relationship between sodium channel hypersensitivity and impaired nerve cell function
Scientific Reports (2025)
-
Identification of biomarkers for the diagnosis in colorectal polyps and metabolic dysfunction-associated steatohepatitis (MASH) by bioinformatics analysis and machine learning
Scientific Reports (2024)
-
Divergent syntheses of complex Veratrum alkaloids
Nature Communications (2024)
-
Pre-clinical safety and therapeutic efficacy of a plant-based alkaloid in a human colon cancer xenograft model
Cell Death Discovery (2022)
-
Human neuronal signaling and communication assays to assess functional neurotoxicity
Archives of Toxicology (2021)
