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4 protocols using α conotoxin mii

1

Pharmacological Modulation of Nicotinic Receptors

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d‐glucose, calcium d‐gluconate, DMSO, ACh, ATP, mecamylamine, NaHCO3, papaine, l‐cysteine, leupeptine, pyruvic acid, NaOH and tyrphostin AG490 (AG 490) were acquired from Sigma‐Aldrich (Taufkirchen, Germany). A‐85380, α‐bungarotoxin, α‐conotoxin PnIA, α‐conotoxin MII, chelerythrine chloride, dihydro‐β‐erythroidine (DHβE), epibatidine, T16Ainh‐A01, Ani 9, CaCCinh‐A01, Chromanol293B, ACV 1 (conotoxin Vc1.1), dantrolene, JTV519 and H‐89 were obtained from Tocris Bioscience (Abingdon, UK). The α‐conotoxin ImI was ordered from Alomone Labs (Jerusalem, Israel). KH2PO4, KCl and MgCl2 were purchased from MERCK (Darmstadt, Germany). NaCl was ordered from Grüssing GmbH (Westoverledingen, Germany). HEPES and BSA were from Carl Roth (Karlsruhe, Germany). Thapsigargin and WP1066 were acquired from Cayman Chemicals (Ann Arbor, MI, USA) and nicotine from Glentham Life Sciences (Corsham, UK). EDTA and Tween 20 were obtained from VWR (Darmstadt, Germany). Sodium pyruvate was purchased from Gibco (Thermo Fisher Scientific, Waltham, MA, USA) and DNase1 from Invitrogen (Thermo Fisher Scientific).
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2

Radioligand Binding Assay Reagents

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[7,8-3H]Dopamine (21.2 Ci/mmol (0.784 TBq/mmol)) and [1-7,8-3H]noradrenaline (12.1 Ci/mmol (0.448 TBq/mmol)) were purchased from Perkin Elmer, Monza, Italy. Nicotine hydrogen tartrate salt, choline, 4-aminopyridine, 6-nitroquipazine, desipramine, beta-amyloid (40-1), horseradish peroxidase-conjugated anti-mouse secondary antibody, Ponceau Red were from Sigma (Sigma-Aldrich, St Louis, MO, USA); Monoclonal antibody 6E10 was from Covance ImmunoTechnologies, Dedham, MA; 5-iodo-A-85380, dihydro-β-erythroidine, α-conotoxin MII, oxotremorine sesquifumarate, desformylflustrabromine hydrochloride, galantamine hydrobromide, SR 165845, beta-amyloid (1-40) were from Tocris (Tocris Bioscience, Bristol, UK).
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Varenicline and Ethanol Pharmacology

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Ethanol (95% v v−1; Gold Shield Chemical Co., Hayward, CA, USA) was diluted with dH2O to 20% solutions. Varenicline (6,7,8,9-tetrahydro-6,10-methanol-6H pyrazino[2,3-h][3]benzazepine tartrate) was generously provided by Pfizer Global Research and Development (Groton, CT, USA). For systemic injections, Varenicline (1.0 mg kg−1, s.c.) dissolved in saline (0.9%) or vehicle (saline, s.c.) was administered in a volume of 1 mL kg−1, and ethanol (20%, 2.5 g kg−1, i.p.) or vehicle (dH2O, i.p.) was administered during in vivo microdialysis. For microinfusion experiments, Varenicline (0.5, 1 and 2 µg) was dissolved in 10% potassium phosphate buffer/90% isotonic saline solution, and thereafter, the pH was adjusted to 7.4 with NaOH (1 N). Vehicle infusions consisted of 10% potassium phosphate buffer/90% isotonic saline. The broad-spectrum β2* nAChR-antagonist dihydro-β-erythroidine (DHβE, 100 nM) and the α6* nAChR-selective antagonist α-conotoxin MII [α-CTXMII at 30 nM concentrations has no effect at non-α6/α3 nAChRs (Cartier et al., 1996 (link))] were purchased from Tocris Bioscience (Bristol, UK) and dissolved in saline. Drug and molecular target nomenclature accords with the British Journal of Pharmacology's Concise Guide to Pharmacology (Alexander et al., 2013a (link)).
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4

Radioligands and Pharmacological Tools for Neuroscience Research

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[7,8-3H] Dopamine (21.2 Ci/mmol (0.784 TBq/mmol)) and [1-7,8-3H] Noradrenaline (12.1 Ci/mmol (0.448 TBq/mmol)) were purchased from Perkin Elmer, Monza, Italy. Nicotine hydrogen tartrate salt was from Sigma (Sigma-Aldrich, St. Louis, MO, USA). 5-iodo-A-85380, α-conotoxin MII, oxotremorine, desformylflustrabromine hydrochloride, beta-amyloid (1–40) were from Tocris (Tocris Bioscience, Bristol, UK). AnnexinV conjugate PE was from Miltenyi Biotec GmbH (order n° 130-118-363) and CalceinAM was from Biotium (catalog n° 30026). KLVFF (segment Aβ 16-20) and VFLKF peptide were purchased from Aurogene s.r.l. (Rome, Italy)
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