Skip to main content
SpringerLink
Log in

Inhibition by anaesthetics of 14C-guanidinium flux through the voltage-gated sodium channel and the cation channel of the 5-HT3 receptor of N1E-115 neuroblastoma cells

  • Published:
Naunyn-Schmiedeberg's Archives of Pharmacology Aims and scope Submit manuscript

Summary

The influence of local and general anaesthetics on cation influx through the fast, voltage-dependent sodium channel and the 5-HT3 receptor cation channel was studied in N1E-115 mouse neuroblastoma cells by measuring 2-min influx of the organic cation 14C-guanidinium induced by either veratridine (1 mmol/l) or 5-HT (100 μmol/l).

The veratridine-induced influx of 14C-guanidinium was potentiated by scorpion toxin and inhibited by tetrodotoxin. The 5-HT-induced 14C-guanidinium influx was not affected by tetrodotoxin but it was inhibited by nanomolar concentrations of the selective 5-HT3 receptor antagonists ondansetron and ICS 205–930; at high micromolar concentrations these compounds also inhibited the veratridine-induced influx of 14C-guanidinium. The 14C-guanidinium influx through both channels was inhibited by local and general anaesthetics. The rank order of potency for inhibition of veratridine-induced influx by local anaesthetics was tetracaine > bupivacaine > cocaine > lidocaine > procaine and that for inhibition of the 5-HT3 receptor channel was tetracaine > bupivacaine > cocaine > procaine > lidocaine. With the exception of procaine and cocaine, which were equipotent at both channels, the local anaesthetics were 4.4-fold (lidocaine) to 25-fold (tetracaine) more potent at the fast sodium channel than at the 5-HT3 receptor channel. The rank order of potency for general anaesthetics was propofol > etomidate = alfaxalone = ketamine > thiopental = methohexital at the fast sodium channel, and propofol ≥ etomidate > alfaxalone = methohexital > thiopental > ketamine at the 5-HT3 receptor channel. With the exception of ketamine, which was about equipotent at both channels, the general anaesthetics were between 2.2 to 8.1-fold more potent at the 5-HT3 receptor channel than at the fast sodium channel. Propofol and alfaxalone non-competitively inhibited the 5-HT-induced influx of 14C-guanidinium. The pIC50 values of local and general anaesthetics for inhibition of veratridine-induced 14C-guanidinium influx were correlated with the lipophilicity (log P values) of these compounds, whereas no correlation was found between the corresponding paramaters for the 5-HT-induced influx.

In conclusion, the results indicate that not only the fast Na+ channel but also the 5-HT3 receptor channel is a target for local and general anaesthetics. The relative high inhibitory potencies of the general anaesthetics which were not correlated with log P values argue in favour of a specific interaction of these agents with the 5-HT3 receptor channel.

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

Similar content being viewed by others

References

  • Amano T, Richelson E, Nirenberg PG (1972) Neurotransmitter synthesis by neuroblastoma clones. Proc Natl Acad Sci USA 69:258–263

    Google Scholar 

  • Barann M, Bönisch H (1992) Inhibition by anaesthetics and steroids of 5-HT3 receptor-mediated influx of 14C-guanidinium ions in neuroblastoma cells. Naunyn-Schmiedeberg's Arch Pharmacol 345 [Suppl]:R112

    Google Scholar 

  • Barann M, Graupner J, Göthert M, Bönisch H (1991) Properties of the 5-HT3-receptor on N1E-115 neuroblastoma cells studied by guanidinium ion influx. Naunyn-Schmiedeberg's Arch Pharmacol 344 [Suppl]:R36

    Google Scholar 

  • Benoit E, Carratu MR, Dubois JM, Mitolo-Chieppa D, Preziosi P (1987) Electrophysiological studies of the effects of the general anaesthetic etomidate on frog myelinated nerve fibre. Br J Pharmacol 90:7–14

    Google Scholar 

  • Benoit E, Carratu MR, Mitolo-Chieppa D (1988) Interactions between molecules of a steroid anaesthetic (alphaxalone) and ionic channels of nodal membrane in voltage-clamped myelinated nerve fibre. Br J Pharmacol 94:635–646

    Google Scholar 

  • Blaustein MP (1968) Barbiturates block sodium and potassium conductance increase in voltage-clamped lobster axons. J Gen Physiol 51:293–307

    Google Scholar 

  • Bönisch H, Barann M, Graupner J, Göthert M (1993) Characterization of 5-HT3 receptors of N1E-115 neuroblastoma cells by use of the influx of the organic cation [14C]-guanidinium. Br J Pharmacol (in press)

  • Burch PG, Stansky DR (1983) The role of metabolism and protein binding in thiopental anesthesia. Anesthesiology 58:146–152

    Google Scholar 

  • Carlon GC, Khan RC, Goldener PL, Howlands WS, Turnbull A (1978) Long-term infusion of sodium-thiopental. Crit Care Med 6:311–316

    Google Scholar 

  • Catterall WA (1980) Neurotoxins that act on voltage-sensitive sodium channels in excitable membranes. Ann Rev Pharmacol Toxicol 20:15–43

    Google Scholar 

  • Concas A, Santoro G, Mascia MP, Serra M, Sanna E, Biggio G (1990) The general anesthetics propofol enhances the function of γ-aminobutyric acid-coupled chloride channel in the rat cerebral cortex. J Neurochem 55:2135–2138

    Google Scholar 

  • Cottrell GA, Lambert JJ, Peters JA (1987) Modulation of GABAA receptor activity by alphaxalone. Br J Pharmacol 90:491–500

    Google Scholar 

  • Derkach V, Surprenant A, North RA (1989) 5-HT3 receptors are membrane ion channels. Nature 339:706–709

    Google Scholar 

  • De Ruiter G, Popescu DT, De Boer AG, Smeekens JB, Breimer DD (1981) Pharmacokinetics of etomidate in surgical patients. Arch Int Pharmacodyn 249:180–188

    Google Scholar 

  • Dilger JP, Firestone LL (1990) More models described for molecular “target” of anaesthetics and alcohols. Trends Pharmacol Sci 11:431–432

    Google Scholar 

  • Dluzewski AR, Halsey MJ, Simmonds AC (1983) Membrane interactions with general and local anaesthetics: a review of molecular hypotheses of anaesthesia. Mol Aspects Med 6:459–573

    Google Scholar 

  • El-Maghrabi EA, Eckenhoff RG, Shuman H (1992) Saturable binding of halothane to rat brain synaptosomes. Proc Natl Acad Sci USA 89:4329–4332

    Google Scholar 

  • Forman SA, Miller KW (1989) Molecular sites of anesthetic action in postsynaptic nicotinic membranes. Trends Pharmacol Sci 10:447–452

    Google Scholar 

  • Fozard JR (1990) Agonists and antagonists of 5-HT3 receptors. In: Saxena PR, Wallis DI, Wouters W, Bevan P (eds) Cardiovascular pharmacology of 5-hydroxytryptamine; Kluwer Academic Publishers, Dordrecht, Boston, London pp 101–115

    Google Scholar 

  • Franks NP, Lieb WR (1984) Do general anaesthetics act by competitive binding to specific receptors. Nature 310:599–601

    Google Scholar 

  • Frenkel C, Urban BW (1991) Human brain sodium channels as one of the molecular target sites for the new intravenous anaesthetic propofol (2,6-diisopropyl-phenol). Eur J Pharmacol — Mol Pharmacol Sect 208:75–79

    Google Scholar 

  • Gepts E, Camu F, Cockshott ID, Douglas EJ (1985) Disposition of propofol administered as zero-order intravenous infusions in humans. Anesth Analg 64:1256–1263

    Google Scholar 

  • Glanville TN, Spence MW, Cook HW, Palmer FBStC (1988) Sodium channel activation does not alter lipid metabolism in cultured neuroblastoma cells. Neurochem Res 13:1015–1021

    Google Scholar 

  • Göthert M (1978) Modification of catecholamine release by anaesthetics and alcohols. In: Paton DM (ed) The release of catecholamines from adrenergic neurons. Pergamon Press, Oxford, New York, pp 241–261

    Google Scholar 

  • Göthert M, Dorn W, Loewenstein I (1976 a) Inhibition of catecholamine release from the adrenal medulla by halothane. Naunyn-Schmiedeberg's Arch Pharmacol 294:239–249

    Google Scholar 

  • Göthert M, Kennerknecht E, Thielecke G (1976 b) Inhibition of receptor-mediated noradrenaline release from the sympathetic nerves of the isolated rabbit heart by anaesthetics and alcohols on proportion to their hydrophobic property. Naunyn-Schmiedeberg's Arch Pharmacol 292:145–152

    Google Scholar 

  • Göthert M, Dührsen U, Rieckesmann JM (1979) Ethanol, anaesthetics and other lipophilic drugs preferentially inhibit 5-hydroxytryptamine and acetylcholine-induced noradrenaline release from sympathetic nerves. Arch Int Pharmacodyn Ther 242:196–209

    Google Scholar 

  • Haky JE, Young AM (1984) Evaluation of a simple HPLC correlation method for the estimation of the octanol-water partition coefficient of organic compounds. J Liq Chromatogr 7:675–689

    Google Scholar 

  • Hales TG, Lambert JJ (1991) The actions of propofol on inhibitory amino acid receptors of bovine adrenomedullary chromaffin cells and rodent central neurones. Br J Pharmacol 104:619–628

    Google Scholar 

  • Honerjäger P, Frelin C, Lazdunski M (1982) Actions, interactions and apparent affinities of various ceveratrum alkaloids at sodium channels of cultured neuroblastoma and cardiac cells. Naunyn-Schmiedeberg's Arch Pharmacol 321:123–129

    Google Scholar 

  • Hoyer D, Neijt HC (1988) Identification of serotonin 5-HT3 recognition sites in membranes of N1E-115 neuroblastoma cells by radioligand binding. Mol Pharmacol 33:303–309

    Google Scholar 

  • Jacques Y, Fosset M, Lazdunski M (1978) Molecular properties of the action potential Na+ ionophore in neuroblastoma cells. Interactions with neurotoxins. J Biol Chem 253:7383–7392

    Google Scholar 

  • Johnston RR, Miller RD, Way WL (1974) The interaction of ketamine with d-tubocurarine, pancuronium, and succinylcholine in man. Anesth Analg 53:496–501

    Google Scholar 

  • Kilpatrick GJ, Bunce KT, Tyers MB (1990) 5-HT3 receptors. Medic Res Rev 10:441–475

    Google Scholar 

  • Kojda G, Klaus W, Werner G, Fricke U (1991) The influence of 3-ester side chain variation on the cardiovascular profile of nitrendipine in porcine isolated trabeculae and coronary arteries. Naunyn-Schmiedeberg's Arch Pharmacol 344:488–494

    Google Scholar 

  • Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193:265–275

    Google Scholar 

  • Maricq AV, Peterson AS, Brake AJ, Myers RM, Julius D (1991) Primary structure and functional expression of the 5-HT3 receptor, a serotonin-gated ion channel. Science 254:432–437

    Google Scholar 

  • McNeal ET, Lewandowski GA, Daly JW, Creveling CR (1985) [3H]Batrachotoxinin 20 -benzoate binding to voltage-sensitive sodium channels: A rapid and quantitive assay for local anaesthetic activity in a variety of drugs. J Med Chem 28:381–388

    Google Scholar 

  • Meyer H (1899) Zur Theorie der Alkoholnarkose. Erste Mittheilung. Welche Eigenschaft der Anästhetica bedingt ihre narkotische Wirkung? Arch Exp Pathol Pharmakol 42:109–118

    Google Scholar 

  • Miller KW, Braswell LM, Firestone LL, Dodson BA, Forman SA (1986) General anaesthetics act both specifically and nonspecifically on acetylcholine receptors. In: Roth SH, Miller KW (eds) Molecular and cellular mechanisms of anaesthetics. Plenum, New York, pp 125–137

    Google Scholar 

  • Neijt HC, TeDuits IJ, Vijverberg HPM (1988) Pharmacological characterization of serotonin 5-HT3 receptor-mediated electrical response in cultured mouse neuroblastoma cells. Neuropharmacology 27:301–307

    Google Scholar 

  • Neijt HC, Plomp JJ, Vijverberg HPM (1989) Kinetics of the membrane current mediated by serotonin 5-HT3 receptors in cultured mouse neuroblastoma cells. J Physiol (Lond) 411:257–269

    Google Scholar 

  • Olsen RW (1982) Drug interactions at the GABA receptor-ionophore complex. Ann Rev Pharmacol Toxicol 122:245–277

    Google Scholar 

  • Overton E (1901) Studien über die Narkose zugleich ein Beitrag zur allgemeinen Pharmakologie. G. Fischer, Jena

    Google Scholar 

  • Peters JA, Usherwood PNR (1983) 5-Hydroxytryptamine responses of murine neuroblastoma cells. Ions and putative antagonists. Br J Pharmacol 180: 523P

  • Proctor WR, Mynlieff M, Dunwiddie TV (1986) Facilitatory action of etomidate and pentobarbital on recurrent inhibition in rat hippocampal pyramidal neurons. J Neurosci 6:3161–3168

    Google Scholar 

  • Reiser G, Günther A, Hamprecht B (1983) Blockade by neurotransmitter antagonists of veratridine-activated ion channels in neuronal cell lines. J Neurochem 40:493–502

    Google Scholar 

  • Reith MEA (1990) 14C-guanidinium ion influx into Na+-channel preparations from mouse cerebral cortex. Eur J Pharmacol 188:33–41

    Google Scholar 

  • Richards CD (1983) Actions of general anaesthetics on synaptic transmission in the CNS. Br J Anaesth 55:201–207

    Google Scholar 

  • Scholtysik G, Imoto Y, Yatani A, Brown AM (1988) 5-Hydroxytryptamine antagonist ICS 205–930 blocks cardiac potassium, sodium and calcium currents. J Pharmacol Exp Ther 245:773–778

    Google Scholar 

  • Schüttler J, Stoeckel H, Schwilden H (1985) Pharmacokinetic and pharmacodynamic modelling of propofol (“Diprivan”) in voltunteers and surgical patients. Postgrad Med J 61:53–54

    Google Scholar 

  • Sear JW, Prys-Roberts C (1979) Plasma concentrations of alphaxalone during continuous infusion with Althesin. Br J Anaesth 51:861–865

    Google Scholar 

  • Shafer A, Doze VA, Shafer SL, White PF (1988) Pharmacokinetics and pharmacodynamics of propofol infusions during general anaesthesia. Anesthesiology 69:348–353

    Google Scholar 

  • Takara H, Wada A, Arita A, Sumikawa K, Izumi F (1986) Ketamine inhibits 45Ca influx and catecholamine secretion by inhibiting 22Na influx in cultured bovine adrenal medullary cells. Eur J Pharmacol 125:217–224

    Google Scholar 

  • Tas PWL, Kress HG, Koschel K (1987) General anaesthetics can competitively interfere with sensitive membrane proteins. Proc Natl Acad Sci USA 84:5972–5975

    Google Scholar 

  • Todd MM, Brummond JC, Hoi Sang U (1984) The chemodynamic consequences of high-dose methohexital anesthesia in humans. Anesthesiology 61:495–501

    Google Scholar 

  • Ueda I, Kamaya H (1973) Kinetic and thermodynamic aspects of the mechanism of general anaesthetics in a model system of firefly luminescence in vitro. Anesthesiology 38:425–436

    Google Scholar 

  • Van Hamme MJ, Ghoneim MM, Ambre JJ (1978) Pharmacokinetics of etomidate, a new intravenous anesthetic. Anesthesiology 49:274–277

    Google Scholar 

  • Wachtel RE, Wegrzynowicz ES (1992) Kinetics of nicotinic acetylcholine ion channels in the presence of intravenous anaesthetics and induction agents. Br J Pharmacol 106:623–627

    Google Scholar 

  • White PF, Way WL, Trevor AJ (1982) Ketamine: Its pharmacology and therapeutic uses. Anesthesiology 56:119–136

    Google Scholar 

  • Wieber J, Gugler R, Hengstmann JH, Dengler HJ (1975) Pharmacokinetics of ketamine in man. Anaesthesist 24:260–263

    Google Scholar 

  • Yamamura T, Harada K, Okamura A, Kemmotsu O (1990) Is the site of action of ketamine anesthesia the N-methyl-D-aspartate receptor? Anesthesiology 72:704–710

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Institut für Pharmakologie and Toxikologie der Universität Bonn, Reuterstrasse 2b, W-5300, Bonn 1, Federal Republic of Germany

    Martin Barann, Manfred Göthert, Klaus Fink & Heinz Bönisch

Authors
  1. Martin Barann
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Manfred Göthert
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Klaus Fink
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Heinz Bönisch
    View author publications

    You can also search for this author in PubMed Google Scholar

Additional information

Correspondence to: H. Bonisch at the above address

Rights and permissions

Reprints and permissions

About this article

Cite this article

Barann, M., Göthert, M., Fink, K. et al. Inhibition by anaesthetics of 14C-guanidinium flux through the voltage-gated sodium channel and the cation channel of the 5-HT3 receptor of N1E-115 neuroblastoma cells. Naunyn-Schmiedeberg's Arch Pharmacol 347, 125–132 (1993). https://doi.org/10.1007/BF00169256

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00169256

Key words

Search

Navigation