Mianserin hydrochloride

Manufactured by Merck Group

Sourced in Poland, United States

Mianserin hydrochloride is a chemical compound used in laboratory research. It is a heterocyclic compound with the molecular formula C18H21N3·HCl. This product is intended for use in scientific investigations and research applications.

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Mianserin (4 citations) Hydrochlorides (2 citations)

8 protocols using mianserin hydrochloride

Pharmacological Compounds Procurement Protocol

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Sertraline hydrochloride, U0126, and SB202190 were purchased from Wako Pure Chemicals Industries (Osaka, Japan). Fluoxetine hydrochloride was purchased from LKT Labs (St. Paul, MN, United States). Imipramine hydrochloride and mianserin hydrochloride were purchased from Sigma–Aldrich. Fluvoxamine maleate and SP600125 were purchased from Tokyo Chemical Industries (Tokyo, Japan). Other common laboratory reagents were also obtained from commercial sources.

Kinboshi M., Mukai T., Nagao Y., Matsuba Y., Tsuji Y., Tanaka S., Tokudome K., Shimizu S., Ito H., Ikeda A., Inanobe A., Kurachi Y., Inoue S, & Ohno Y. (2017). Inhibition of Inwardly Rectifying Potassium (Kir) 4.1 Channels Facilitates Brain-Derived Neurotrophic Factor (BDNF) Expression in Astrocytes. Frontiers in Molecular Neuroscience, 10, 408.

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Preparation of Drosophila Food with Pharmacological Compounds

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Rapamycin (LC Laboratories) was dissolved in ethanol and added to SYA food at a final concentration of 400 µM [17] (link). Tolbutamide (Santa Cruz Biotechnology) was dissolved in ethanol and added to SYA food at a final concentration of 1.35 mg/ml [74] (link). Cycloheximide and mianserin hydrochloride (Sigma) were dissolved in water and added to SYA food at final concentrations of 17 mM [75] (link) and 0.2 mg/ml [32] (link), respectively. METH hydrochloride (Sigma) was dissolved in ethanol and added to SYA food at a final concentration of 1 mg/ml [3] (link),[4] (link). 3IY was added directly to SYA food at a final concentration of 5 mg/ml [3] (link). An overview of pharmacological sites of action can be found in Figure S8.

Metaxakis A., Tain L.S., Grönke S., Hendrich O., Hinze Y., Birras U, & Partridge L. (2014). Lowered Insulin Signalling Ameliorates Age-Related Sleep Fragmentation in Drosophila. PLoS Biology, 12(4), e1001824.

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Caffeine and Antidepressant Interaction Study

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Caffeine (5 mg/kg; Sigma-Aldrich, Poznań, Poland) and mianserin hydrochloride (10 mg/kg; Sigma-Aldrich) were dissolved in saline (0.9% sodium chloride (NaCl)). Agomelatine (20 mg/kg; Sigma-Aldrich) was suspended in a 1% aqueous solution of Tween 80 (POCH, Gliwice, Poland). The doses and treatment schedules were selected based on our previous experiments (Poleszak et al. 2016 (link)). The mice were randomly assigned to groups that received the solution of Caffeine or saline intraperitoneally (i.p.), twice a day (at 8 a.m. and 8 p.m.), for 14 days. Both groups were further divided into subgroups that received on the 15th day Caffeine or saline 40 min before behavioral testing and the solutions/suspensions of antidepressants or saline 60 min before behavioral testing. All solutions/suspensions were prepared fresh before administration and were administered at a volume of 0.1 ml per 10 g of body weight. The number of animals used per group was 10 in the FST, TST, and pharmacokinetic studies and 8 in the locomotor activity test.

Szopa A., Poleszak E., Doboszewska U., Herbet M., Świąder K., Wyska E., Serefko A., Wlaź A., Korga A., Ostrowska M., Juś P., Jedynak S., Dudka J, & Wlaź P. (2018). Withdrawal of caffeine after its chronic administration modifies the antidepressant-like activity of atypical antidepressants in mice. Changes in cortical expression of Comt, Slc6a15 and Adora1 genes. Psychopharmacology, 235(8), 2423-2434.

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Radioligand Binding Assay Protocol

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All purchased compounds were >98% pure according to the manufacturers. Coelentrazine H (Thermo Scientific, Waltham, MA) was dissolved in ethanol as a stock solution (5 mM). Serotonin (5-HT) hydrochloride (Acros Organics, ThermoFischer Scientific, Pittsburgh, PA), d-luciferin (Gold Biotechnology, St. Louis, MO), and mianserin hydrochloride (Sigma Aldrich, St. Louis, MO) were dissolved in dimethyl sulfoxide (DMSO) (10 mM) and prepared fresh daily. [³H]-Mesulergine (84.7 Ci/mmol) and [³H]-ketanserin (47.3 Ci/mmol) were purchased from PerkinElmer Life Sciences (Waltham, MA).

Felsing D.E., Anastasio N.C., Miszkiel J.M., Gilbertson S.R., Allen J.A, & Cunningham K.A. (2018). Biophysical validation of serotonin 5-HT2A and 5-HT2C receptor interaction. PLoS ONE, 13(8), e0203137.

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Serotonin and Biogenic Amine Pharmacology

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Most of the chemical compounds were obtained from Sigma-Aldrich, Inc (St. Louis, MO) including serotonin hydrochloride (≥98%), selected biogenic amines (dopamine hydrochloride (≥98%), (±)-octopamine hydrochloride (≥95%), tyramine hydrochloride (≥98%), and histamine (97%)), agonists (5-methoxytryptamine (97%), α-methyl-5-HT, 1,(3-chlorophenyl)piperazine hydrochloride (99%), tryptamine hydrochloride (99%), quipazine maleate salt (≥98%)), antagonists (methiothepin mesylate salt (≥98%), yohimbine hydrochloride (≥98%), cyproheptadine hydrochloride sesquihydrate (99%), mianserin hydrochloride (≥98%), ketanserin tartrate salt (97%), and selective serotonin reuptake inhibitor, fluoxetine hydrochloride (≥98%). Stock solutions of the compounds were prepared with dimethyl sulfoxide (DMSO) for the in vitro experiments and ultrapure H₂O (Invitrogen) for the in vivo experiments, and stored at −20 °C. 4-chloro-DL-chlorophenylalanine ethyl ester hydrochloride (98%) (PCPA) was purchased from Alfa Aesar (Ward Hill, MA) and prepared in 10% sucrose immediately before use.

Ngai M., Shoue D.A., Loh Z, & McDowell M.A. (2019). The pharmacological and functional characterization of the serotonergic system in Anopheles gambiae and Aedes aegypti: influences on flight and blood-feeding behavior. Scientific Reports, 9, 4421.

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Innate Immune Response Analysis

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PRR ligands were obtained from Invivogen and used at the following working concentrations: Pam₃CSK₄ (1 μg/mL), HKLM (10⁸ cells/mL), poly(I:C) (25 μg/mL), LPS (1 μg/mL), flagellin (1 μg/mL), FSL‐1 (1 μg/mL), imiquimod (10 μg/mL), ssRNA40 (10 μg/mL), ssRNA41 (10 μg/mL), ODN2006 (3 μM), poly(I:C)/LyoVec (10 μg/mL), and MDP (10 μg/mL). Chloroquine and mianserin hydrochloride were obtained from Sigma. Influenza A/PR/8/34 stock (400 hemagglutination units (HAU)/mL) was kindly provided by Prof. John McCauley (NIMR, London). CD80, CD83, CD86 antibodies for flow cytometry were purchased from eBioscience. Polyclonal rabbit anti‐MxA antibody used for immunoblot and flow cytometry was purchased from Abnova (H00004149‐D01P). The Rabbit (DA1E) mAb IgG XP^® antibody (CST, #3900) was used as matching isotype control for flow cytometry. Pre‐miRs and anti‐miRs were obtained from Ambion (pre‐miR negative control #1: AM17110, anti‐miR negative control #1: AM17010, pre‐miR‐650: PM11602, anti‐miR‐650: AM11602).

Pichulik T., Khatamzas E., Liu X., Brain O., Delmiro Garcia M., Leslie A., Danis B., Mayer A., Baban D., Ragoussis J., Weber A.N, & Simmons A. (2015). Pattern recognition receptor mediated downregulation of microRNA‐650 fine‐tunes MxA expression in dendritic cells infected with influenza A virus. European Journal of Immunology, 46(1), 167-177.

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5-HT Radioligand Binding Assay

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Serotonin (5-HT, Acros Organics, Thermo Fisher Scientific, Fair Lawn, NY) was dissolved in 1x HBSS (Cellgro; Invitrogen, Manassas, VA). Benzyl 2-acetamido-2-deoxy-α-D-galactopyranoside (GalNac-O-Bn, Sigma-Aldrich, St. Louis, MO) was dissolved in serum free Dulbecco’s modified Eagle’s medium. All purchased compounds for the radioligand binding assay were >98% pure according to the manufacturers. Mianserin hydrochloride (Sigma Aldrich, St. Louis, MO) was dissolved in dimethyl sulfoxide (DMSO) (10 mM) and prepared fresh daily. [³H]-Mesulergine (83.0 Ci/mmol) was purchased from PerkinElmer Life Sciences (Waltham, MA).

Land M.A., Chapman H.L., Davis-Reyes B.D., Felsing D.E., Allen J.A., Moeller F.G., Elferink L.A., Cunningham K.A, & Anastasio N.C. (2019). Serotonin 5-HT2C Receptor Cys23Ser Single Nucleotide Polymorphism Associates with Receptor Function and Localization In Vitro. Scientific Reports, 9, 16737.

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Ligand Binding Assay Reagents

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Tyramine hydrochloride, (±)-octopamine hydrochloride, dopamine hydrochloride, naphazoline hydrochloride, forskolin, chlorpromazine hydrochloride, phentolamine hydrochloride, and mianserin hydrochloride were all purchased from Sigma-Aldrich (St. Louis, MO, USA). To culture HEK-293 cells, DMEM/F12 medium, fetal bovine serum (FBS), fungizone, penicillin, and streptomycin were purchased from Gibco Cell Culture at Life Technologies (Grand Island, NY, USA). TransIT-LT1 Transfection Reagent was obtained from Mirus Bio LLC (Madison, WI, USA).

Li H.M., Jiang H.B., Gui S.H., Liu X.Q., Liu H., Lu X.P., Smagghe G, & Wang J.J. (2016). Characterization of a β-Adrenergic-Like Octopamine Receptor in the Oriental Fruit Fly, Bactrocera dorsalis (Hendel). International Journal of Molecular Sciences, 17(10), 1577.

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