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12 protocols using citalopram hydrobromide

1

Comprehensive Pharmacology Reagents Protocol

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We purchased common salts and 1-(3-Chlorophenyl) biguanide hydrochloride (mCPBG); gabazine; 1,2-bis(2-aminophenoxy) ethane-N,N,N,N-tetraacetic acid (BAPTA); SCH50911; 6,7-dinitroquinoxaline-2,3-dione (DNQX); tetrodotoxin (TTX); strychnine; SB200646 hydrochloride (SB200646); ritanserin (RIT); ondansetron hydrochloride dihydrate (OND); and 5-HT hydrochloride (5-HT) from Sigma-Aldrich Chemical Company (St Louis, MO, USA). Citalopram hydrobromide, WAY100635, and 1-(3-Chlorophenyl) piperazine hydrochloride (mCPP) from Tocris Bioscience (Ellisville, MO, USA).
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2

Pharmacological Modulation of Neurotransmitter Signaling

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We purchased 1-(3-Chlorophenyl) biguanide hydrochloride (mCPBG), gabazine, DL-2-amino-5-phosphono-valeric acid (DL-APV), 1,2-bis(2-aminophenoxy)ethane-N,N,N,N-tetraacetic acid (BAPTA), SCH50911, 6, 7-dinitroquinoxaline-2, 3-dione (DNQX), tetrodotoxin (TTX), strychnine, SB-200646 hydrochloride (SB200646), ritanserin (RIT), ondansetron hydrochloride dihydrate (OND) and serotonin hydrochloride (5-HT), 2-Aminoethyl diphenylborinate (2-APB) and other common salts from Sigma-Aldrich Chemical Company (St Louis, MO, USA); and Citalopram hydrobromide, WAY100635 and 1-(3-Chlorophenyl) piperazine hydrochloride (mCPP) from Tocris Bioscience (Ellisville, MO, USA).
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3

Pharmacological Modulation of Monoaminergic Systems

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Ketamine and xylazine were obtained from Midwest Veterinary Supply (Lakeville, MN). Meloxicam was obtained from Norbrook Laboratories (Overland Park, KS). Atomoxetine hydrochloride (1044469) and fluoxetine hydrochloride (F132) were obtained from Sigma-Aldrich (St. Louis, MO). Citalopram hydrobromide (1427), reboxetine mesylate (1982), N-(2-Chloroethyl)-N-ethyl-2-bromobenzylamine hydrochloride (DSP-4,2958), prazosin hydrochloride (0623), (S)-duloxetine hydrochloride (4798), propranolol hydrochloride (0624), and (R)-(−)-phenylephrine hydrochloride (2838) were obtained from Tocris Biosciences (Minneapolis, MN). Fluoxetine, citalopram, prazosin, and reboxetine were dissolved in 1% DMSO. DSP-4, phenylephrine, atomoxetine, propranolol, and duloxetine were dissolved in saline (0.9% NaCl).
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4

Preparation of Psychoactive Drug Solutions

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Yohimbine hydrochloride (Tocris, UK) was dissolved in distilled water. Citalopram hydrobromide (Tocris, UK), buspirone hydrochloride (Sigma, UK) amphetamine (Sigma, UK) and alpha-flupenthixol (Sigma, UK) were dissolved in 0.9 % saline. FG7142 (β-carboline-3-carboxylic acid N-methylamide) and diazepam (both Sigma, UK) were dissolved in 10 % dimethyl sulphoxide (DMSO; BDH Laboratory Supplies, UK), 20 % cremophor EL (Sigma, UK) and 70 % of 0.9 % saline. All drugs were administered at a final volume of 1 ml/kg.
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5

Intracerebral Drug Infusion Protocol

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Citalopram hydrobromide (Tocris), reboxetine mesylate (Tocris), paroxetine hydrochloride hemihydrate (Sigma), imipramine hydrochloride (Sigma), and methysergide (Sigma) were freshly dissolved into saline (NaCl 0.9%) before use. Compounds were dissolved in a final volume of 10mL/kg. α-FMHis (synthesized at Abbott Laboratories, Chicago, IL) was injected i.c.v. at the dose of 5 µg dissolved in 5 µL of saline. All doses were calculated as mg/kg of the free base. Control animals received saline. In reverse dialysis experiments, drugs were diluted in the perfusing Ringer’s solution. All other reagents and solvents were of high performance liquid chromatography (HPLC) grade or the highest grade available (Sigma).
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6

Pharmacological Modulation of Serotonergic Pathways

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Chloral hydrate, 8-OH-DPAT hydrobromide, and WAY100635 maleate were obtained from Sigma-Aldrich. Citalopram hydrobromide and tertiapin-Q were from Tocris Bioscience. tertiapin-Q was prepared in ACSF. Chloral hydrate, 8-OH-DPAT, WAY100635, and citalopram were prepared in 0.9% saline.
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7

Drug Infusion and Injection Techniques

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Veratridine, dihydrokainic acid (DHK), 2,3-dioxo-6-nitro-1,2,3,4-tetrahydrobenzo[f]quinoxaline-7-sulfonamide disodium salt (NBQX), (S)-α-Amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (s-AMPA) and citalopram hydrobromide were purchased from Tocris (Bristol, UK). 4-chloro-dl-phenylalanine methyl ester hydrochloride (pCPA) was purchased from Sigma-Aldrich (Tres Cantos, Spain).
For intracerebral microinfusion, Veratridine was dissolved in 12.5% dimethyl sulfoxide in artificial cerebrospinal fluid and pH adjusted to 6.5; DHK and s-AMPA were dissolved in PBS 10 × as previously reported;29 NBQX and citalopram were dissolved in artificial cerebrospinal fluid. The microinfusion volume was 0.5 μl per side in all cases. pCPA was administered intraperitoneally, was daily prepared by dissolving in saline and pH was adjusted to 6.0 with 0.1 m NaOH.30 (link)
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8

Pharmacological Selectivity of Antidepressants

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Citalopram hydrobromide, fluoxetine hydrochloride, and duloxetine hydrochloride (Tocris; Bristol, United Kingdom) were each dissolved in sterile saline at a concentration of 2 mg/ml and injected at a volume of 10 μl/g body weight for a dose of 20 mg/kg. This dose of citalopram has been shown to reduce responding for a conditioned reinforcer in wild-type but not serotonin transporter knockout mice, suggesting pharmacological selectivity (Browne and Fletcher, 2016 (link)). All saline and drug injections were administered i.p. Animals remained in their homecage for 30 minutes after injections before being placed in chambers for testing.
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9

Pharmacological Exploration of 5-MeO-DMT Mechanisms

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5-Methoxy-N,N-dimethyltryptamine (5-MeO-DMT), risperidone (RIS) and WAY-100635 maleate were from Sigma/RBI (Natick, MA). Haloperidol (HAL) was from Laboratorios Esteve (Barcelona, Spain). Citalopram hydrobromide was from Tocris (Bristol, UK). Doses are expressed as free bases. All drugs were dissolved in saline and injected subcutaneously (s.c.). For the assessment of local effects in mPFC, 5-MeO-DMT was dissolved in the artificial cerebrospinal fluid (aCSF) used to perfuse the microdialysis probes (see below).
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10

Modulation of Stress Responses in Mice

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C57Bl/6 mice were treated intraperitoneally (i.p.) with the NK1 receptor antagonist CP99994 (25-50 mg/kg, Tocris; dissolved in saline) 30 minutes prior to measurements. As a negative control, one group of animals received the same amount of saline. As a positive control in acute stress tests, one animal group was treated intraperitoneally (i.p.) with citalopram hydrobromide, a selective serotonin reuptake inhibitor (10 mg/kg, Tocris; dissolved in saline) 30 minutes prior to measurements.
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