Specific details of swimming assays of wild type, mutant and drug-treated animals are provided in Supplementary Methods . Imipramine HCl was obtained from Sigma (St. Louis, MO). Swip assays were performed as described in Hardaway and Hardie et al. Concentration response curves were fit by nonlinear regression methods as implemented in Prism 6.0 (GraphPad, Inc, La Jolla, CA). Average frequency values were analyzed by 2-way ANOVA with Dunnet’s post tests of each time point in the drug condition to the corresponding basal time point.
Imipramine hcl
Manufactured by Merck Group
Sourced in United States
Imipramine HCl is a pharmaceutical compound that functions as a tricyclic antidepressant. It is commonly used as a reference standard in analytical testing and research applications.
Automatically generated - may contain errors
Lab products found in correlation
Fluoxetine hcl, by Merck Group (2 mentions)
Prism 6, by GraphPad (1 mentions)
Ethylene diamine tetraacetic acid (edta), by Merck Group (1 mentions)
Milli q water purification system, by Merck Group (1 mentions)
Fluvoxamine maleate, by Merck Group (1 mentions)
Citalopram hbr, by Merck Group (1 mentions)
Desipramine hcl, by Merck Group (1 mentions)
Clomipramine hcl, by Merck Group (1 mentions)
Apomorphine, by Merck Group (1 mentions)
Xylazine, by Bayer (1 mentions)
Ketamine, by Ceva (1 mentions)
Amitriptyline hcl, by Merck Group (1 mentions)
Alprazolam, by Merck Group (1 mentions)
Chlordiazepoxide hcl, by Merck Group (1 mentions)
Nitroblue tetrazolium, by Merck Group (1 mentions)
1 1 3 3 tetramethoxypropane, by Merck Group (1 mentions)
Reduced glutathione, by Merck Group (1 mentions)
Bovine serum albumin (bsa), by Merck Group (1 mentions)
Ethylenediaminetetraacetic acid, by Merck Group (1 mentions)
Xanthine oxidase, by Merck Group (1 mentions)
8 protocols using imipramine hcl
1
Comprehensive Swimming Assay Protocol
2
Detailed Protocol for Analytical Quantification
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ZnPT, PT, MSP, EDTA, imipramine HCl (an internal standard for MSP analysis), and d3-5-Nitro-5′-hydroxy-indirubin-3′-oxime (d3-AGM130; an internal standard for ZnPT and PT analyses) were purchased from Sigma-Aldrich Chemical Corporation (Milwaukee, WI, USA). Water was purified with a Milli-Q water purification system (Millipore, Bedford, MA, USA). All other chemicals and reagents were of analytical grade and used without further purification.
3
Imipramine Administration in Rats
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Experimentally naïve male Sprague-Dawley rats (Harlan, Italy) aged about 10 weeks and weighing 300–350 g at the beginning of the experiment were used as subjects. The animals were housed in groups of two-three per cage in controlled environmental conditions (temperature 22–24° C; humidity 50–60%; light on at 08:00, off at 20:00), with free access to food and water. Imipramine HCl (Sigma, St Louis, USA) was dissolved in distilled water and injected intra-peritoneally (i.p.) in a volume of 1 ml/kg. Vehicle treatment consisted in a 1 ml/kg distilled water i.p. administration.
4
Phytochemical and Antidepressant Evaluations
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All reagents used in the determination of the total phenolic and flavonoid contents were purchased from Sigma-Aldrich Chemical Co. (USA). Spectroscopy measurements were performed on a UV-Vis Shimadzu Multispect-1501 spectrophotometer (Kyoto, Japan). In animal experiments, ethanolic extract of each plant was suspended in distilled water using Tween® 80 (1%). Fluoxetine HCL (Sigma-Aldrich, USA) and imipramine HCL (Sigma-Aldrich, USA) were both dissolved in normal saline. The plant extracts, fluoxetine, imipramine, and vehicle were injected intraperitoneally (10 mL/kg, i.p.) 30 min before each experiment.
5
Radioligand Binding Assays for Monoamine Transporters
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All SSRIs (alaproclate-HCl, citalopram-HBr, fluvoxamine-maleate, fluoxetine-HCl, norfluoxetine-HCl, paroxetine-HCl, sertraline-HCl) and other monoamine transporter inhibitors (imipramine-HCl, desipramine-HCl, clomipramine-HCl, maprotiline-HCl) were obtained from Sigma-Aldrich (St. Louis, USA). [125I]MIBG (specific activity ≈ 0.65 TBq/mmol) was purchased from Chelatec (Saint-Herblain, France). [3H]Serotonin (5-hydroxytryptamine creatinine sulfate, specific activity ≈ 3.84 TBq/mmol) was purchased from PerkinElmer.
6
Unilateral 6-OHDA Lesion Model
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To protect noradrenergic neurons, animals were administered with imipramine HCl (15 mg/kg, ip, Sigma) 15 minutes before surgery. They were subsequently anesthetized with ketamine (Ceva, 75 mg/kg) and xylazine (Bayer, 10 mg/kg) and placed into a stereotaxic frame (David Kopf Instrument). 6-OHDA was injected into the right ascending medial forebrain bundle at the following coordinates (in mm) relative to bregma and surface of the dura, AP = -3.5, ML = -1.5, DV = -8.7 [27 ]. Each rat received one injection of 6-OHDA (4 μg/μl) over a period of 5 minutes (0.5 μl/min) for a total of 10 μg per rat. Animals were monitored for 3 weeks to ensure full recovery and habituation to the environment and experimenters. On day 21 after surgery, all rats were challenged with a small subcutaneous dose of apomorphine (Sigma, 0.05 mg/kg). Rats showing more than 90 contraversive rotations (360°) over a 45-minute recording period were included in the study. It has been previously demonstrated that rats meeting this criterion have a unilateral loss of dopaminergic neurons and a unilateral depletion of striatal dopamine of over 95% [19 (link)].
7
Pharmacological Characterization of LY2940094
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LY2940094 was synthesized as described by Eli Lilly and Company (Toledo et al. 2014 ). The compound was dissolved in 20% captisol®, a cyclodextrin derivative (Cydex, Inc., Lenexa, KS) in 25 mmol/L phosphate buffer (pH2) and dosed orally 60 min prior to testing. Imipramine HCl, amitriptyline HCl, and chlordiazepoxide HCl were obtained from Sigma‐Aldrich. They were dissolved in 0.9% NaCl and injected i.p., 30 min prior to testing. Paroxetine HCl and fluoxetine HCl were synthesized at Eli Lilly and Company, dissolved in 0.9% NaCl, and dosed i.p., 30 min prior to testing. MTEP and MPEP (Tocris) were dissolved in water and dosed i.p. Alprazolam (Sigma‐Aldrich) was suspended in 10% hydroxypropyl beta‐cyclodextrin. Compounds were dosed in the forms noted in a volume of 1 or 2 mL/kg for rat studies and 10 mL/kg for mouse studies.
8
Analytical Characterization of Synthetic SAC
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SAC was synthetized as described previously [21 (link)], and was compared with a SAC standard based on melting point, 1H nuclear magnetic resonance spectroscopy, and infrared spectroscopy [21 (link)]. GR, NADPH, xanthine, xanthine oxidase, nitroblue tetrazolium (NBT), bovine serum albumin (BSA), ethylenediaminetetraacetic acid (EDTA), reduced glutathione (GSH), glutathione disulfide (GSSG), H2O2, sodium diethyldithyocarbamate trihydrate (DDC), trichloroacetic acid (TCA), dihydroethidium (DHE), 1,1,3,3-tetramethoxypropane, and imipramine-HCl were from Sigma-Aldrich (St. Louis, MO, USA). Thiobarbituric acid (TBA), 2′,7′-dichlorofluorescein diacetate (DCFH-DA), and 2′,7′-dichlorofluorescein (DCF) were from Merck (Darmstadt, Germany). All other reagent-grade chemicals were commercially available.
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