Methyllycaconitine citrate

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Methyllycaconitine citrate is a chemical compound used in research applications. It is a selective antagonist of the α7 nicotinic acetylcholine receptor. This product is intended for laboratory use only.

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Methyllycaconitine citrate (MLA; (10 citations)

7 protocols using methyllycaconitine citrate

Pharmacological Modulation of Neuronal Signaling

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For in vitro experiments, SR 95531 hydrobromide (gabazine 5 μM, Tocris Bioscience, UK, Ref. 1262), NMDA (10 μM, Tocris Bioscience, UK, Ref. 0114), 2,3-dihydroxy-6-nitro-7-sulfamoyl-benzo[f]quinoxaline-2,3-dione (10 µM, NIH generous gift), GABA (10 μM, Sigma-Aldrich), isoguvacine (10 μM, Sigma-Aldrich, Ref. G002), DL -2-Amino-5-phosphonovaleric acid (40 µM, Sigma-Aldrich, Ref. A5282), and bumetanide (10 μM, Sigma-Aldrich, Ref. B3023) were directly added to the perfusion solutions. For ca experiments, slices were treated with bumetanide for 40 min before and during recordings. Cocktail of nicotinic receptor antagonists included mecamylamine hydrochloride (10 μM, Tocris Bioscience, Ref. 2843/10), methyllycaconitine citrate (0.1 μM, Tocris Bioscience, Ref. 1029/5), and dihydro-β-erythroidine hydrobromide (10 μM, Tocris Bioscience, Ref. 2349/10). For in vivo experiments, bumetanide pretreatment (3 mg kg⁻¹) was given to mice in drinking water during 5 weeks.

Lozovaya N., Eftekhari S., Cloarec R., Gouty-Colomer L.A., Dufour A., Riffault B., Billon-Grand M., Pons-Bennaceur A., Oumar N., Burnashev N., Ben-Ari Y, & Hammond C. (2018). GABAergic inhibition in dual-transmission cholinergic and GABAergic striatal interneurons is abolished in Parkinson disease. Nature Communications, 9, 1422.

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Neuroprotective Effects of Selective α7nAChR Modulation

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The following drugs were used in the study: PNU-282987 (an α7nAChR agonist, 1 mg/kg/day, Abcam, ab120558) (25 (link)–27 (link)), Methyllycaconitine citrate (MLA, an α7nAChR antagonist, 1 mg/kg/day, Tocris Bioscience, 1029) (28 (link)–30 (link)) and 3-MA (an autophagy inhibitor, 10 mg/kg/day, MedChemExpress, HY-19312) (23 (link)). All drugs were injected intraperitoneally. Based on salt weight and concentration, compounds were dissolved in 0.9% saline, the injection volume is 1 ml/kg body weight. The first dose was administered 1 hour after ICH induction, followed by daily doses at 24-hour intervals. Control animals were injected with saline.

Su Y., Zhang W., Zhang R., Yuan Q., Wu R., Liu X., Wuri J., Li R, & Yan T. (2022). Activation of Cholinergic Anti-Inflammatory Pathway Ameliorates Cerebral and Cardiac Dysfunction After Intracerebral Hemorrhage Through Autophagy. Frontiers in Immunology, 13, 870174.

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Pharmacological Effects of MDMA and M100907

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Racemic MDMA [(±)-3,4-methylenedioxymethamphetamine hydrochloride] and M100907 were kindly provided by the National Institute of Drug Abuse (Baltimore, MD, USA). Ketanserin tartrate and methyllycaconitine citrate were purchased from TOCRIS (Ellisville, MO, USA). Hexamethonium bromide, midazolam hydrochloride, and WAY100,635 maleate were purchased from Sigma (St. Louis, MO, USA). Drug dosages (mg/kg) were expressed as the weight of the salt and injected at a constant volume of 1 mL/kg (dissolved in 0.9% NaCl). MDMA and ketanserin were injected intraperitoneally, and other drugs were administered subcutaneously.

Tao R., Shokry I.M., Callanan J.J., Adams H.D, & Ma Z. (2014). Mechanisms and environmental factors that underlying the intensification of 3,4-methylenedioxymethamphetamine (MDMA, Ecstasy)-induced serotonin syndrome in rats. Psychopharmacology, 232(7), 1245-1260.

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Radioligand Binding Assay for Neuronal Nicotinic Receptors

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All salts and reagents were ACS grade and purchased from Fisher Scientifics (www.fishersci.com), Sigma-Aldrich (www.sigmaaldrich.com), and Tocris Biosciences (www.tocris.com). [³H]-methyllycaconitine (100 Ci/mmol in ethanol/water) from American Radiolabeled Chemicals (www.arcincusa.com); methyllycaconitine citrate from Tocris Biosciences (www.tocris.com); D-amphetamine hemisulfate was purchase from Sigma-Aldrich; L-amphetamine hydrochloride from Lipomed Inc (www.lipomed.com); and (±)-1-methyl-2- pyridin-3-ylethylamine from Broadpharm (www.broadpharm.com). Antibodies for tyrosine hydroxylase and the dopamine transporter were purchased from EMD Millipore (www.emdmillipore.com). Xenopus laevis oocytes were obtained from EcoCyte Bioscience (www.ecocyte-us.com).

Garton D.R., Ross S.G., Maldonado-Hernández R., Quick M., Lasalde-Dominicci J.A, & Lizardi-Ortiz J.E. (2018). Amphetamine Enantiomers Inhibit Homomeric α7 Nicotinic Receptor through a Competitive Mechanism and within the Intoxication Levels in Humans. Neuropharmacology, 144, 172-183.

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Neuronal Calcium Signaling Assay

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Triton X-100, (-)-nicotine hydrogen tartrate and mecamylamine hydrochloride, were purchased from Sigma-Aldrich (Poole, Dorset, UK); B27, l-glutamine, antibiotics, fluo-3 AM, fura-2 AM, and pluronic f127 were obtained from Life Technologies (Paisley, UK); sazetidine-A dihydrochloride, tetrodotoxin citrate, methyllycaconitine citrate and 5-iodo-A85380 dihydrochloride were purchased from Tocris Bioscience (Avonmouth, UK); PNU-120596 and PNU-282987 were provided by Pfizer Inc. USA; general reagents were purchased from Fisher Scientific (Loughborough, UK).

Brown J.L, & Wonnacott S. (2014). Sazetidine-A Activates and Desensitizes Native α7 Nicotinic Acetylcholine Receptors. Neurochemical Research, 40(10), 2047-2054.

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Neurotransmitter Receptor Pharmacology

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Acetylcholine chloride and methyllycaconitine citrate were obtained from Tocris (Minneapolis, MN, United States). Strychnine hydrochloride, dimethoxystrychnine sulfate hydrate (brucine), atropine sulfate monohydrate (-)-nicotine hydrogen tartrate, sodium chloride, potassium chloride, calcium chloride dihydrate, magnesium chloride hexahydrate, 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES), actinomycin-D, α-Bgtx from Bungurus multicinctus, α-cobratoxin from Naja kaouthia, were obtained from Sigma-Aldrich (St. Louis, MO, United States).

Hone A.J, & McIntosh J.M. (2022). Alkaloid ligands enable function of homomeric human α10 nicotinic acetylcholine receptors. Frontiers in Pharmacology, 13, 981760.

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Antibodies and Reagents for Cell Signaling

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Antibodies directed against caspase 3 and -actin were obtained from Cell Signaling Technology (Beverly, Massachusetts, USA). Nrf2 antibody was purchased from Santa Cruz Biotechnology (Dallas, Texas, USA). HO-1 antibody was obtained from ABCAM (Cambridge, UK). H 2 O 2 was obtained from Merck (Darmstadt, Germany). Electrochemiluminescence (ECL) kit was obtained from Amersham Bioscience (Piscataway, New Jersey, USA). Polyvinylidene fluoride (PVDF) was from Millipore (Billerica, Massachusetts, USA). Tropisetron and methyllycaconitine citrate (MLA) were purchased from Tocris Bioscience (Bristol, BS11 0QL, UK). All the other reagents, unless otherwise stated, were from Sigma Aldrich (St Louis, Missouri, USA).

Khalifeh S., Fakhfouri G., Mehr S.E., Mousavizadeh K., Mousavizadeh K., Dehpour A.R., Khodagholi F., Kazmi S, & Rahimian R. (2015). Beyond the 5-HT3 receptors: A role for α7nACh receptors in neuroprotective aspects of tropisetron. Human & experimental toxicology, 34(9).

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