tetrodotoxin  (Alomone Labs)

Journal: PLoS ONE

Article Title: Neuroligin-1 Loss Is Associated with Reduced Tenacity of Excitatory Synapses

doi: 10.1371/journal.pone.0042314

Figure Lengend Snippet: Activity dependence of AMPAR redistribution dynamics. A ) Distribution of range over mean values for SEpH:GluA2 puncta in the presence of CNQX (10 µM), AP-5 (50 µM) and TTX (1 µM) (Nlgn-1 KO: 10 neurons, 481 synapses; WT: 10 neurons, 514 synapses). B ) SI decay rates in the presence of the aforementioned pharmacological agents.

Article Snippet: Pharmacological manipulations Reagents were procured from the following sources: CNQX (6-cyano-7 nitroquinoxaline-2,3-dione) from Tocris Bioscience; AP-5 (2-amino-5-phosphonopentanoic acid) from Sigma-Aldrich; TTX (tetrodotoxin) from Alomone Labs.

Techniques: Activity Assay

Journal: PLoS ONE

Article Title: The Voltage-Gated Sodium Channel Nav1.8 Is Expressed in Human Sperm

doi: 10.1371/journal.pone.0076084

Figure Lengend Snippet: Effects of veratridine on human sperm motility in the presence of tetrodotoxin, A-803467 or ab-66743. The effects of veratridine (10 μM) after different incubation times were analyzed in the presence of (A) the VGSC inhibitor tetrodotoxin (TTX) (10 nM) (B) TTX (10 μM), (C) the selective Na v 1.8 antagonist A-803467 (10 μM), (D) the Na v 1.8 antibody ab-66743 (dilution 1:50) or the corresponding solvent. Bars are means with SEM of 6-8 different experiments and represent percentage changes in progressive motility (grade A+B sperm) relative to the value observed at the same time in the respective solvent-treated paired controls. * P

Article Snippet: In parallel experiments, the effect of veratridine or its solvent was investigated in aliquots pretreated during the last 15 min of capacitation with the specific Na v 1.8 antagonist A-803467 (Sigma) (1 μM) [ ] or during capacitation (2 h) with TTX (Alomone Labs, Jerusalem, Israel)) (0.01 or 10 μM), the Na v 1.8 antibody ab-66743 (dilution 1:50) or the corresponding solvent.

Techniques: Incubation

Journal: bioRxiv

Article Title: Neuroepithelial progenitors generate and propagate non-neuronal action potentials across the spinal cord

doi: 10.1101/2020.05.23.111955

Figure Lengend Snippet: Floor-plate biphasic action potentials are triggered by the activation of nicotinic acetylcholine receptors in response to acetylcholine released by motoneurons. a, Example of recurrent spontaneous floor-plate action potentials blocked after addition of the nicotinic acetylcholine receptor (nAChR) antagonists: Mecamylamine (50 μM) and d-Tubocurarine (5 μM). b, Example of current-clamp recording showing floor-plate action potential evoked by electrical stimulation in control condition (black trace) and after addition of the nAChR antagonists (red trace). c, Example of current-clamp recording showing floor-plate action potential evoked in control condition (black trace) and after addition of antagonists against ionotropic receptor for GABA (Gabazine 3 μM) and glutamate (DL-APV 200 μM and CNQX 20 μM). d, Example of current-clamp recording showing that floor-plate action potential can be evoked by local application of 30 μM acetylcholine (left trace), even in the presence of TTX (1 μM) and antagonists to AMPA/Kainate glutamate receptor (CNQX 10 μM), NMDA glutamate receptor (DL-APV 200 μM), GABA A receptor (Gabazine 3 μM) and glycine receptor (strychnine 1 μM) (center trace). Floor-plate action potential evoked by acetylcholine were blocked by the addition of nAChR antagonists (right trace). Note that the addition of TTX inhibited the fast component of the biphasic action potential (see Supplementary Figure 2 ). e, Confocal image of a coronal section from a ChAT:ChR2-YFP mouse embryo at E12.5 showing the expression of Channelrhodopsin2-YFP fusion protein (in green) in cholinergic motoneurons located in the ventro-dorsal horns and labelled with the vesicular acetylcholine transporter vAChT (in red). All cell nuclei were labelled using DAPI (in blue). f, Example of current-clamp recording from a ChAT:ChR2-YFP + motoneuron showing an action potential triggered by the opening of Channelrhodopsin 2 in response to blue light stimulation (470 nm). g, Example of current-clamp recording from a floor-plate cell recorded in a ChAT:ChR2-YFP + fetal spinal cord showing how blue light stimulation could evoke a slow cholinergic depolarization and trigger a biphasic action potential that were blocked by the addition of nAChR antagonists.

Article Snippet: The following pharmacological agents were used: Acetylcholine chloride (30 μM, Sigma-Aldrich, Germany), TTX (1 μM, Alomone Labs, Israel), gabazine (3 μM, Tocris, USA), CNQX Disodium (20 μM, Tocris, USA), DL-APV (200 μM, Tocris, USA), Mecamylamine hydrochloride (100 μM, Tocris, USA), D-turbocurarine chloride (10 μM,Tocris Bioscience, Minneapolis, MN, USA), 18-β glycyrrhetinic acid (50 μM, Sigma-Aldrich, Germany) and TTA-P2 (3μM, Alomone, Israel).

Techniques: Activation Assay, Expressing

Journal: Journal of neuroendocrinology

Article Title: Gonadal steroids differentially modulate the actions of orphanin FQ/nociceptin at a physiologically relevant circuit controlling female sexual receptivity

doi: 10.1111/jne.12148

Figure Lengend Snippet: Membrane current traces that illustrate the OFQ/N-induced outward currents recorded in identified POMC neurones from OVX rats either 30 hr after the initial vehicle or EB priming and four hr after the second vehicle or progesterone (P4) treatment (A-C), or 48 hr after a high dose of EB (50 μg) that elicits sexual receptivity much like that seen in EB-primed, P4-treated animals (D). These recordings were performed in the presence of 1 μM TTX to ensure a direct postsynaptic effect. For each example, OFQ/N application commenced as signified by the left-hand side of the OFQ/N application timeline box located above each set of traces. I/V relationships were routinely generated prior to, and in the presence of, OFQ/N. This necessitated switching between gap-free and episodic protocols, which precluded the acquisition of a single trace showing both the outward current and its wash-out. The time necessary to conduct the second I/V and then switch back to the gap-free recording of washout took, at most, 90 sec. The two panels, both of equal dimensions, were aligned along their respective X-axes. The time required for complete OFQ/N washout was variable, and in some instances, it took as long as 20 min to be achieved. This is why the time scales for the two halves of the recording are different.

Article Snippet: OFQ/N, the voltage-gated Na+ channel blocker tetrodotoxin (TTX) with citrate (Alomone Labs, Jerusalem, Israel), the GABAA receptor antagonist 6-imino-3-(4-methoxyphenyl)-1(6H)- pyridazinebutanoic acid hydrobromide (SR 95531) and the α-amino-3-hydroxyl-5-methyl-4-isoxazole- propionate (AMPA) receptor antagonist 2,3-dioxo-6-nitro-1,2,3,4-tetrahydrobenzo[f]quinoxaline-7- sulfonamide (NBQX) were dissolved in UltraPure H2 0 to stock concentrations of 1 mM, 1 mM, 10 mM and 10 mM, respectively.

Techniques: Generated, Size-exclusion Chromatography