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Tetrodotoxin

Manufactured by Alomone
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Sourced in Israel, Australia, United Kingdom, United States

Tetrodotoxin is a highly potent neurotoxin that acts by blocking sodium channels, preventing the generation and propagation of action potentials in nerve and muscle cells. It is commonly found in various marine organisms, including pufferfish, blue-ringed octopus, and certain species of newts.

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Lab products found in correlation

Picrotoxin, by Merck Group (3 mentions) Cdcl2, by Merck Group (3 mentions) Strychnine, by Merck Group (3 mentions) Picrotoxin, by Bio-Techne (2 mentions) Xe991, by Bio-Techne (2 mentions) Iberiotoxin, by Alomone (2 mentions) 15nh4cl, by Merck Group (2 mentions) Isopropyl β d thiogalactopyranoside iptg and streptomycin, by Thermo Fisher Scientific (2 mentions) 13c6 glucose, by Merck Group (2 mentions) Hplc grade acetonitrile, by RCI Labscan (2 mentions) Phrixotoxin 1, by Alomone (2 mentions) Guangtoxin 1e, by Alomone (2 mentions) Xe 991, by Alomone (2 mentions) Tolbutamide, by Merck Group (2 mentions) 2 arachidonylglycerol, by Bio-Techne (2 mentions) 1 2 4 dichlorophenyl 5 4 iodophenyl 4 methyl n 4 morpholinyl 1h pyrazole 3 carboxamide am281, by Merck Group (2 mentions) Bf150 86 7, by Sutter Instruments (2 mentions) 8 cyclopentyl 1 3 dipropylxanthine dpcpx, by Bio-Techne (2 mentions) 6 n n diethyl d β γ dibromomethylene atp trisodium salt arl 67156 trisodium salt, by Bio-Techne (2 mentions) Metamorph 7, by Molecular Devices (1 mentions)

Spelling variants of this product

Tetrodotoxin (TTX) (42 citations) Tetrodotoxin citrate (6 citations)

37 protocols using tetrodotoxin

1

Tetrodotoxin Inhibits Myofiber Contractions

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For action potential-blocking experiments, the selective NaV channels blocker tetrodotoxin (final concentration 1 µM, Alomone Labs) was applied to the co-culture medium. Response in myofibers was recorded by video microscopy at 2 frames per second and analyzed using Metamorph 7.1 (Molecular Devices). Contractions of myofibers were monitored by following displacement of the plasma membrane of one point over time using the Track points plug in.
Vilmont V., Cadot B., Ouanounou G, & Gomes E.R. (2016). A system for studying mechanisms of neuromuscular junction development and maintenance. Development (Cambridge, England), 143(13), 2464-2477.
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2

Assessing Synaptic Plasticity in Mature Hippocampal Neurons

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Mature hippocampal neurons (7 × 104 in culture on 15 mm coverslips) were incubated with 2 × 107 ctrl-EVs or Aβ-EVs in 330 µl, or vehicle, for 1 h. To record miniature excitatory post-synaptic currents (mEPSCs), the voltage-gated Na+-channel blocker tetrodotoxin (TTX, 0.5 μM, Alomone Labs) and the GABA-A receptor antagonist picrotoxin (100 μM, Merck) were added to the bath solution, along with strychnine (1 μM, Merck), used to avoid glycine receptor activation. In order to investigate synaptic plasticity, after 12 min baseline recording in standard bath solution, Mg2+-free bath solution was perfused for 1 min before applying the same solution containing glycine (Gly, 200 μM, Merck) for 3 min. Subsequently, perfusion was resumed with standard Mg2+-containing bath solution and recording continued for 40 min. Potentiation magnitude was measured as the average response between the 28th and the 40th min after glycine. See also Supplementary material.
Gabrielli M., Prada I., Joshi P., Falcicchia C., D’Arrigo G., Rutigliano G., Battocchio E., Zenatelli R., Tozzi F., Radeghieri A., Arancio O., Origlia N, & Verderio C. (2022). Microglial large extracellular vesicles propagate early synaptic dysfunction in Alzheimer’s disease. Brain, 145(8), 2849-2868.
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3

Tetrodotoxin and GBP Pharmacological Assay

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Tetrodotoxin was purchased from Alomone Laboratories (Jerusalem, Israel), and GBP was purchased from Tocris Bioscience (Bristol, United Kingdom). Drugs were stored at 1000× final concentration and diluted in oocyte bath solution or aCSF prior to application. All data were pooled and represent means ± standard error of the mean (SEM). Data sets were compared using Student’s t-test unless stated (GraphPad Prism 7; GraphPad Software, La Jolla, CA, United States). An F test was applied to determine equality of variance before parametric statistical analysis was applied (GraphPad Prism). Differences were deemed statistically significant when p < 0.05.
Tae H.S., Smith K.M., Phillips A.M., Boyle K.A., Li M., Forster I.C., Hatch R.J., Richardson R., Hughes D.I., Graham B.A., Petrou S, & Reid C.A. (2017). Gabapentin Modulates HCN4 Channel Voltage-Dependence. Frontiers in Pharmacology, 8, 554.
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4

Molecular signaling mechanisms in neurons

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Reagents: picrotoxin and strychnine were purchased from Sigma (St. Louis, MO, USA) and were dissolved in ethanol (96%, Sigma) and water, respectively. Tetrodotoxin (TTX; Alomone Labs, Jerusalem, Israel) was dissolved in water.
Primary Antibodies: anti-pTrkAY490 (Cell Signalling, Danvers, MA, USA, 9141), anti-Trk (Santa Cruz, Santa Cruz, CA, USA, sc7268), anti-pShcY317 (Cell Signalling, 2434), anti-ShcC (Cell Signalling, 2431), anti-pPLC-γ Y783 (Cell Signalling, 2821), anti PLC-γ (Cell Signalling, 14008), anti-pMAPKY202/Y204 (Cell Signalling, 9101), anti-MAPK (Cell Signalling, 9102), anti-pPI3KY458/Y199 (Cell Signalling, 4228), anti-pAKTS473 (Cell Signalling, 4051), anti-AKT (11E7) (Cell Signalling, 4685), anti-pCREBS133 (Millipore, Temecula, CA, USA, 06-519), anti-BDNF (Millipore, MABN110), anti-c-Fos (Cell Signalling, Rb Mab 2250S), anti-ChAT (Millipore, AB144P), and anti-NeuN (ABCAM, city, state abbrev if USA, country ab177487), anti-MAP2 (Cell Signalling, 4542; Millipore, MAB3418). Secondary antibodies: anti mouse-HRP and anti rabbit-HRP (PerkinElmer, Waltham, MA, USA); Donkey anti mouse-Alexa546, donkey anti rabbit-Alexa488 (Life Technologies, Carlsbad, CA, USA).
Triaca V., Fico E., Sposato V., Caioli S., Ciotti M.T., Zona C., Mercanti D., La Mendola D., Satriano C., Rizzarelli E., Tirassa P, & Calissano P. (2020). hNGF Peptides Elicit the NGF-TrkA Signalling Pathway in Cholinergic Neurons and Retain Full Neurotrophic Activity in the DRG Assay. Biomolecules, 10(2), 216.
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5

Neurotransmitter Receptor Modulation

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Tetrodotoxin was purchased from Alomone Labs (Jerusalem, Israel), bicuculline methiodide and picrotoxin were purchased from Tocris (Minneapolis, MN, USA), d-cycloserine (DCS), escitalopram and desipramine were purchased from Sigma-–Aldrich (St. Louis, MO, USA). Ro-25-6981 was purchased from Abcam chemicals (Cambridge, MA, USA).
Gupta S.C., Yadav R., Pavuluri R., Morley B.J., Stairs D.J, & Dravid S.M. (2015). Essential role of GluD1 in dendritic spine development and GluN2B to GluN2A NMDAR subunit switch in the cortex and hippocampus reveals ability of GluN2B inhibition in correcting hyperconnectivity. Neuropharmacology, 93, 274-284.
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6

Electrophysiological Recording of Neuronal Cells

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A sucrose solution was used during Vibratome slicing (in mM): 2 KCl, 1 MgCl2–6H2O, NaH2PO4, 10 HEPES, 10 glucose, 208 sucrose, 26 NaHCO3, 2 MgSO4–7H2O, and 1 CaCl2. For harvesting and electrophysiological recordings, a standard artificial cerebrospinal fluid was used (in mM): 124 NaCl, 5 KCl, 1.44 NaH2PO4, 5 HEPES, 10 glucose, 26 NaHCO3, 2 MgSO4–7H2O, and 2 CaCl2. Tetrodotoxin (1 μM, Alomone Labs, Jerusalem, Israel) was added to the bath for all of the recordings except when measuring the rheobase. Tetraethylammonium (TEA, 5 mM, Sigma–Aldrich, St. Louis, MO, USA), a non-specific K+ channel blocker, and XE 991 (40 μM, Tocris, Bristol, UK), a KCNQ channel antagonist, were also used.
Stincic T.L., Bosch M.A., Hunker A.C., Juarez B., Connors A.M., Zweifel L.S., Rønnekleiv O.K, & Kelly M.J. (2021). CRISPR knockdown of Kcnq3 attenuates the M-current and increases excitability of NPY/AgRP neurons to alter energy balance. Molecular Metabolism, 49, 101218.
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7

Synthesis and Characterization of Gambierol

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Gambierol was synthesized as described by Fuwa et al. in 2002 [7 (link)] and had a purity >97%. The synthetic gambierol was spectroscopically (NMR 13C and 1H, MS, IR) identical to natural gambierol. Due to the lipophilic nature of gambierol, stock solutions were prepared in dimethyl sulfoxide (DMSO) and diluted with the external physiological solution. The total DMSO concentration in the test solution did not exceed 0.1%. Tetrodotoxin, apamin, iberiotoxin, and glibenclamide were purchased from Alomone Labs (Jerusalem, Israel), and Latoxan (Portes-lès-Valence, France). All other chemicals, including cell culture media, reagents, tetraethylammonium chloride and 3,4-diaminopyridine, were purchased from Sigma-Aldrich (Saint-Quentin-Fallavier, France).
Benoit E., Schlumberger S., Molgó J., Sasaki M., Fuwa H, & Bournaud R. (2022). Gambierol Blocks a K+ Current Fraction without Affecting Catecholamine Release in Rat Fetal Adrenomedullary Cultured Chromaffin Cells. Toxins, 14(4), 254.
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8

Pharmacological Modulation of Ion Channels

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Drugs were obtained from Sigma-Aldrich unless otherwise stated. CFTRinh172 was purchased from Cedarlane Labs (Burlington, ON, CA), tetrodotoxin was obtained from Alomone labs (Jerusalem, Israel), and lidocaine hydrochloride spray was acquired from Odan Laboratories LTD (Montreal, Canada). Stock solutions of CFTRinh172, lidocaine, atropine, bumetanide, niflumic acid, and L-703606 were dissolved in DMSO. The final concentration of DMSO was less than 0.1%. tetrodotoxin was directly dissolved into purified water.
Luan X., Tam J.S., Belev G., Jagadeeshan S., Murray B., Hassan N., Machen T.E., Chapman L.D, & Ianowski J.P. (2019). Nebulized hypertonic saline triggers nervous system-mediated active liquid secretion in cystic fibrosis swine trachea. Scientific Reports, 9, 540.
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9

Neuronal Solution Perfusion Protocol

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Solutions were applied by gravity from a glass capillary (1.2 mm outer diameter) placed ~1 mm from the neuron under study. Solutions were switched manually using a low dead volume manifold upstream of the glass capillary. CNQX and Gabazine were supplied by Abcam. KB-R7943 Mesylate was supplied by Tocris. Creatine Phosphate was supplied by Santa Cruz Biotech. Cinacalcet was supplied by Toronto Research Chemicals and Tetrodotoxin by Alomone Other reagents were obtained from Sigma-Aldrich.
Martiszus B.J., Tsintsadze T., Chang W, & Smith S.M. (2021). Enhanced excitability of cortical neurons in low-divalent solutions is primarily mediated by altered voltage-dependence of voltage-gated sodium channels. eLife, 10, e67914.
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10

Reagent Procurement for Biological Research

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All chemicals were purchased from Sigma-Aldrich Australia (Castle Hill, NSW, Australia), Sigma-Aldrich USA (St Louis, MO, USA), or Merck Chemicals (Kilsyth, Victoria, Australia) with the exception of isopropyl-β-D-thiogalactopyranoside (IPTG) and streptomycin (Life Technologies, Victoria, Australia), tetrodotoxin (Alomone Labs, Israel), and HPLC-grade acetonitrile (RCI Labscan, Bangkok, Thailand). 13C6-glucose and 15NH4Cl were from Sigma-Aldrich Australia. Recombinant His6-TEV protease (EC 3.4.22.44) was produced in-house using a published protocol62 (link).
Bende N.S., Dziemborowicz S., Mobli M., Herzig V., Gilchrist J., Wagner J., Nicholson G.M., King G.F, & Bosmans F. (2014). A distinct sodium channel voltage-sensor locus determines insect selectivity of the spider toxin Dc1a. Nature communications, 5, 4350.
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