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Tetrodotoxin

Manufactured by Abcam
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Tetrodotoxin is a potent neurotoxin that blocks sodium channels in nerve and muscle cells. It is commonly found in certain marine organisms, such as puffer fish. Tetrodotoxin is used in research applications to study the function of voltage-gated sodium channels and their role in neurophysiology.

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Lab products found in correlation

Bicuculline, by Abcam (3 mentions) 4 aminopyridine, by Abcam (3 mentions) Pclamp 10, by Molecular Devices (3 mentions) Gabazine sr 95531 hydrobromide, by Bio-Techne (2 mentions) Ly341495, by Bio-Techne (2 mentions) D15 peptide pppqvpsrpnrappg, by Bio-Synthesis (2 mentions) Ly379268, by Abcam (2 mentions) Neurobiotin, by Vector Laboratories (2 mentions) Dl ap5, by Abcam (2 mentions) Cgp55845, by Abcam (2 mentions) U73122, by Bio-Techne (2 mentions) Sr141716a, by Bio-Techne (2 mentions) Picrotoxin, by Abcam (2 mentions) Picrotoxin, by Bio-Techne (2 mentions) Nisoldipine, by Merck Group (2 mentions) Strychnine, by Merck Group (2 mentions) Kn 93, by Abcam (2 mentions) Multiclamp 700b, by Molecular Devices (2 mentions) Protease inhibitor mixture cocktail, by Roche (1 mentions) Streptavidin alexa fluor 488, by Thermo Fisher Scientific (1 mentions)

31 protocols using tetrodotoxin

1

Preparation and Storage of Pharmacological Reagents

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MPH was purchased from Sigma-Aldrich (St. Louis, MO), and diluted to a 5 mM stock solution in physiological saline. Prepared solution was stored at −20 °C in 1 ml aliquots. Tetrodotoxin, (TTX) was purchased from Abcam (Cambridge, MA) and stored in 1 mM stock at −20 °C. DNQX was purchased from Tocris (Minneapolis, MN), diluted to 20 mM stock and stored at 4 °C. R,S-CPP was purchased from Abcam (Cambridge, MA), diluted to 10 mM stock and stored at −20 °C. AP5 was purchased from Sigma-Aldrich (St. Louis, MO) and was diluted to a 50 mM stock and stored at −20 °C. Gabazine (SR-95531 hydrobromide) was purchased from Tocris (Minneapolis, MN) and was stored at −20 °C in a 2.5 mM stock.
Urban K.R., Li Y.C., Xing B, & Gao W.J. (2017). A Clinically-Relevant Dose of Methylphenidate Enhances Synaptic Inhibition in the Juvenile Rat Prefrontal Cortex. Journal of reward deficiency syndrome and addiction science, 2(3), 69-77.
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2

Preparation and Storage of Pharmacological Reagents

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MPH was purchased from Sigma-Aldrich (St. Louis, MO), and diluted to a 5 mM stock solution in physiological saline. Prepared solution was stored at −20 °C in 1 ml aliquots. Tetrodotoxin, (TTX) was purchased from Abcam (Cambridge, MA) and stored in 1 mM stock at −20 °C. DNQX was purchased from Tocris (Minneapolis, MN), diluted to 20 mM stock and stored at 4 °C. R,S-CPP was purchased from Abcam (Cambridge, MA), diluted to 10 mM stock and stored at −20 °C. AP5 was purchased from Sigma-Aldrich (St. Louis, MO) and was diluted to a 50 mM stock and stored at −20 °C. Gabazine (SR-95531 hydrobromide) was purchased from Tocris (Minneapolis, MN) and was stored at −20 °C in a 2.5 mM stock.
Urban K.R., Li Y.C., Xing B, & Gao W.J. (2017). A Clinically-Relevant Dose of Methylphenidate Enhances Synaptic Inhibition in the Juvenile Rat Prefrontal Cortex. Journal of reward deficiency syndrome and addiction science, 2(3), 69-77.
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3

Neuromodulatory Compound Procurement and Synthesis

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LY379268 and tetrodotoxin were purchased from Abcam. LY341495 and CNQX were purchased from Tocris. VU0650786 and MRK-8-29 were synthesized in-house. The D15 peptide (PPPQVPSRPNRAPPG) was prepared by Bio-Synthesis.
Joffe M.E., Santiago C.I., Engers J.L., Lindsley C.W, & Conn P.J. (2017). Metabotropic glutamate receptor subtype 3 gates acute stress-induced dysregulation of amygdalo-cortical function. Molecular psychiatry, 24(6), 916-927.
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4

Comprehensive Antibody Panel for Epigenetic Regulation

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Commercial antibodies used were: anti-FLAG (Sigma), anti-CDYL (Sigma, HPA035578), anti-REST (Santa Cruz, sc-25398), anti-EZH2 (Proteintech, 21800-1-AP), anti-HDAC1 (sc-7872), anti-HDAC2 (sc-7899), anti-SIRT1 (sc-15404), anti-FOS (ABclonal, A0236), anti-SET8 (CST, #2996), anti-UTX (sc-79334), anti-KDM5B (sc-67035), anti-CtBP (Proteintech, 10972-1-AP), anti-MeCP2 (ABclonal, A5694), anti-Nav1.6 (Alomone, ASC-009), anti-H3K9me2 (Abcam, ab1220), anti-H3K9me3 (Abcam, ab8898), anti-G9a (Millipore, 09-071), anti-H3K27me3 (Millipore, 07-449), anti-H3K27ac (Abcam, ab4729), anti-H3K4me1 (Abcam, ab8895), anti-H3K4me3 (Abcam, ab8580), anti-GAPDH and anti-Actin (MBL). A CDYL antibody that we generated (peptides antigen: DGFQGESPEKLDPVDQG and IDDRRDQPFDKRLRFSV; B&M) was used for ChIP-seq, western blotting with rat/mouse cell lysates, and immunohistochemistry. Protein A/G beads were from GE Healthcare Biosciences, protease inhibitor mixture cocktail was from Roche Applied Science. Tetrodotoxin, bicuculline, CGP 55845, and DL-AP5 were from Abcam. Neurobiotin was from Vector Laboratories. Streptavidin Alexa Fluor 488 was from Life Technologies. 4,9-TTX was from Focus Biomolecules. All other reagents were purchased from Sigma-Aldrich.
Liu Y., Lai S., Ma W., Ke W., Zhang C., Liu S., Zhang Y., Pei F., Li S., Yi M., Shu Y., Shang Y., Liang J, & Huang Z. (2017). CDYL suppresses epileptogenesis in mice through repression of axonal Nav1.6 sodium channel expression. Nature Communications, 8, 355.
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5

Multielectrode Array Characterization of Neuronal Cultures

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For multielectrode array (MEA) experiments, 1-well MEA plates (60MEA200/30iR-Ti; Multichannel Systems) were used. MEA plates were sterilized by autoclaving and treated with FBS before coating with 0.1% polyethyleneimine and 50 μg/ml laminin. ND1 cells were plated on top of the electrodes in a drop containing 2 × 105 cells in 10 μl of N2 media. Once the cells had adhered to the surface, the wells were slowly flooded with N2 media and plates were returned to the incubator. Media changes were carried out according to the neuronal differentiation protocol described above; half media changes were performed every 4 days once cells were in NB media. Recordings were taken 36–41 days after cells were plated with an MEA 2100 Lite head stage and amplifier with a sample rate of 10,000 Hz. To assess the contribution of glutamatergic signaling and sodium channels to spiking, recordings were repeated in the presence of glutamate antagonist NBQX (10 μM), NMDA antagonist AP-5 (50 μM), and sodium channel blocker tetrodotoxin (1 μM) (all from Abcam). All MEA analysis was done offline with MC Rack software (MultiChannel Systems; RRID:SCR_014955) and NeuroExplorer (RRID:SCR_001818). Spikes were counted when the extracellular recorded signal exceeded 5 standard deviations of the baseline noise.
Zenchak J.R., Palmateer B., Dorka N., Brown T.M., Wagner L.M., Medendorp W.E., Petersen E.D., Prakash M, & Hochgeschwender U. (2018). Bioluminescence-Driven Optogenetic Activation of Transplanted Neural Precursor Cells Improves Motor Deficits in a Parkinson’s Disease Mouse Model. Journal of neuroscience research, 98(3), 458-468.
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6

Pharmacological Modulation of Signaling Pathways

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BIBO3304 and BIIE0246 were purchased from Tocris (Bristol, UK), PSN632408 and OEA were from Cayman Chemical (Ann Arbor, MI), and tetrodotoxin and α-melanocyte–stimulating hormone (α-MSH) were from Abcam Biochemicals (Cambridge, UK). PYY was purchased from Generon (Slough, UK) and VIP from AnaSpec (Freemont, CA). AM1638, AR231453, AR420626, AR435707, AR436352, AR440006, Merck A, TAK875, and Merck V were provided by R.M.J. and T.W.S. All other chemicals were purchased from Sigma-Aldrich (Poole, UK).
Tough I.R., Forbes S., Herzog H., Jones R.M., Schwartz T.W, & Cox H.M. (2018). Bidirectional GPR119 Agonism Requires Peptide YY and Glucose for Activity in Mouse and Human Colon Mucosa. Endocrinology, 159(4), 1704-1717.
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7

Patch-clamp Recordings of Murine Pyramidal Neurons

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Slice recording was performed essentially as described previously [35 (link)]. P14–P18 mouse brains were cut into 350-μm-thick coronal sections. In the current clamp experiments, pyramidal neurons were patched with a glass pipette containing potassium-based intra-cellular solution (ICS) or cesium-based ICS. Miniature postsynaptic currents (mPSCs) were recorded in the presence of 1μM tetrodotoxin (Abcam). Membrane potential was held at −60mV for mEPSCs and 0 mV for mIPSCs. Evoked postsynaptic currents were triggered with a 0.1-ms current injection by a nichrome-wire electrode placed 100–50 μm from the soma of recorded neurons.
Mori T., Kasem E.A., Suzuki-Kouyama E., Cao X., Li X., Kurihara T., Uemura T., Yanagawa T, & Tabuchi K. (2019). Deficiency of calcium/calmodulin-dependent serine protein kinase disrupts the excitatory-inhibitory balance of synapses by down-regulating GluN2B. Molecular Psychiatry, 24(7), 1079-1092.
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8

Pharmacological Modulation of Neural Activity

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Tetrodotoxin (TTX) and picrotoxin were purchased from Abcam, while U73122, SR 141716A, and meta-Chlorophenylpiperazine (mCPP) were purchased from Tocris. Clozapine N-oxide (CNO) was generously provided by Dr. Bryan Roth (University of North Carolina).
Mazzone C.M., Pati D., Michaelides M., DiBerto J., Fox J.H., Tipton G., Anderson C., Duffy K., McKlveen J.M., Hardaway J.A., Magness S.T., Falls W.A., Hammack S.E., McElligott Z.A., Hurd Y.L, & Kash T.L. (2016). Acute engagement of Gq-mediated signaling in the bed nucleus of the stria terminalis induces anxiety-like behavior. Molecular psychiatry, 23(1), 143-153.
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9

Pharmacological Modulation of Neural Activity

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Tetrodotoxin (TTX) and picrotoxin were purchased from Abcam, while U73122, SR 141716A, and meta-Chlorophenylpiperazine (mCPP) were purchased from Tocris. Clozapine N-oxide (CNO) was generously provided by Dr. Bryan Roth (University of North Carolina).
Mazzone C.M., Pati D., Michaelides M., DiBerto J., Fox J.H., Tipton G., Anderson C., Duffy K., McKlveen J.M., Hardaway J.A., Magness S.T., Falls W.A., Hammack S.E., McElligott Z.A., Hurd Y.L, & Kash T.L. (2016). Acute engagement of Gq-mediated signaling in the bed nucleus of the stria terminalis induces anxiety-like behavior. Molecular psychiatry, 23(1), 143-153.
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10

Pharmacological Modulation of Neuronal Excitability

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The following drugs were perfused in aCSF: 100 µM AP5 ([2R]-amino-5-phosphonovaleric acid, an Nmethyl-D-aspartate (NMDA) receptor antagonist, Tocris; IC 50 = ~50 µM), 10 µM CNQX (6-cyano-7nitroquinoxaline-2,3-dione, an AMPA/kainate receptor antagonist, Tocris; IC 50 = 1.5 µM), 2 µM bicuculline (ionotropic g-aminobutyric acid or GABA A receptor antagonist, Sigma-Aldrich; IC 50 = 2 µM), 0.5 µM TTX (tetrodotoxin, a Na + channel blocker, Abcam; IC 50 < 10 nM), and 100 µM 4-AP (4-aminopyridine, a Kv1 channel blocker, Tocris; IC 50 = 147 µM). All drugs were perfused throughout the experimental protocol and washed out for at least 30 min after the end of the protocol.
Lee J., Choi J., & Rah J. (2020). Frequency-dependent gating of feedforward inhibition in thalamofrontal synapses.
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