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.
5 methoxytryptamine
Manufactured by Merck Group
Sourced in United States
5-methoxytryptamine is a chemical compound that can be used for laboratory research purposes. It is a derivative of the neurotransmitter serotonin. The core function of this compound is to serve as a tool for scientific investigation, but its specific intended use should not be extrapolated upon.
Automatically generated - may contain errors
Lab products found in correlation
6 hydroxymelatonin, by Merck Group (3 mentions)
Fluoxetine hydrochloride, by Merck Group (2 mentions)
Cyproheptadine hydrochloride sesquihydrate, by Merck Group (2 mentions)
Melatonin, by Merck Group (2 mentions)
Serotonin hydrochloride, by Merck Group (1 mentions)
Tyramine hydrochloride, by Merck Group (1 mentions)
Tryptamine hydrochloride, by Merck Group (1 mentions)
Yohimbine hydrochloride, by Merck Group (1 mentions)
Mianserin hydrochloride, by Merck Group (1 mentions)
Octopamine hydrochloride, by Merck Group (1 mentions)
Ketanserin tartrate salt, by Merck Group (1 mentions)
Ultrapure h2o, by Thermo Fisher Scientific (1 mentions)
Dopamine hydrochloride, by Merck Group (1 mentions)
Heptadecanoic acid, by Merck Group (1 mentions)
Scopolamine hydrochloride, by Merck Group (1 mentions)
Pentadecanoic acid, by Merck Group (1 mentions)
N acetylaspartylglutamic acid, by Merck Group (1 mentions)
E coli lps, by Merck Group (1 mentions)
N acetylserotonin, by Merck Group (1 mentions)
Pmsg serotropin, by Aska Pharmaceutical (1 mentions)
9 protocols using 5 methoxytryptamine
1
Serotonin and Biogenic Amine Pharmacology
2
Intestinal IgA Binding to Metabolites
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Metabolite-specific ELISA was performed to address the binding of intestinal IgA to purified identified metabolites including pentadecanoic acid (Sigma); 5-methoxytryptamine (Sigma); N-acetylaspartylglutamic acid (Sigma); scopolamine hydrochloride (Sigma); D-maltose (VWR) and heptadecanoic acid (Sigma). E.coli LPS (Sigma) and DNA from E. coli were used as controls. ELISA plates (Milian) were coated with 50μl/well of 10μg/mL antigen overnight (ON) using coating buffer (0.1M NaHCO3) except pentadecanoic acid and heptadecanoic acid coated using 100% ethanol. After blocking (PBS with 2% BSA, RT, 2h), serial diluted intestinal washes containing IgA were added to the plates and incubated for 2 hours. To reveal the IgA binding affinity to metabolites, HRP conjugated anti-IgA antibody (1:100, Sigma) was added to each well and incubated for 45 min. Plates were then washed (4X, PBS containing 0.5% Tween-20) and developing reagent (100 μL per well, 1mg ABTS plus 5μl H2O2/10ml 0.1 M NaH2PO4) was added. Samples then were read at OD415 using an ELISA plate reader.
3
Quantification of Melatonin Derivatives
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The source of chemicals is described previously24 (link). All chemicals were diluted in pure ethanol. AFMK (N1-acetyl-N2-formyl-5-methoxykynuramine) (10005254) was purchased from Cayman Chemical Company (Ann Arbor, MI, USA) (catalog number 10005254), while others were purchased from Sigma-Aldrich (St. Louis, MO): Melatonin (N-acetyl-5-methoxytryptamine) (M5250), 6-hydroxyMelatonin ( 3-(N-acetylaminoethyl)-6-hydroxy-5-methoxyindole) (H0627), N-acetylserotonin (N-acetyl-5-hydroxytryptamine) (A1824), 5-methoxytryptamine ( 2-(5-methoxy-1H-indol-3-yl)ethanamine) (286583).
4
Pharmacological Modulation of Serotonergic Signaling
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We purchased 5-HT, sumatriptan succinate (Sumatriptan), α-methylserotonin maleate salt (MS), 1-(3-chlorophenyl)biguanide hydrochloride (mCPBG), 5-methoxytryptamine (MT), WAY-208466
dihydrochloride (WAY), LP12 hydrochloride hydrate (LP12), cyproheptadine hydrochloride sesquihydrate (Cypro), dolasetron mesylate hydrate (DS), GR113808 (GR), and SB-258719 (SB25) from
Sigma-Aldrich (St. Louis, MO, USA). Pregnant mare serum gonadotropin (PMSG) (Serotropin®) and human chorionic gonadotropin (hCG) (Gonatropin®) were purchased from ASKA
Pharmaceutical (Tokyo, Japan). Other reagent-grade chemicals were purchased from FUJIFILM Wako Pure Chemical Corp. (Osaka, Japan).
dihydrochloride (WAY), LP12 hydrochloride hydrate (LP12), cyproheptadine hydrochloride sesquihydrate (Cypro), dolasetron mesylate hydrate (DS), GR113808 (GR), and SB-258719 (SB25) from
Sigma-Aldrich (St. Louis, MO, USA). Pregnant mare serum gonadotropin (PMSG) (Serotropin®) and human chorionic gonadotropin (hCG) (Gonatropin®) were purchased from ASKA
Pharmaceutical (Tokyo, Japan). Other reagent-grade chemicals were purchased from FUJIFILM Wako Pure Chemical Corp. (Osaka, Japan).
5
Assessing Immune Cell Apoptosis and Oxidative Stress
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The anti-mouse Ly-6A/EA (Sca-1)-PE/Cy7, CD117 (c-kit), APC, biotin-conjugated CD5, CD4, CD8, CD45R/B220, Ly6G/Gr-1, CD11b, Ter-119, and APC/CY7-conjugated streptavidin antibodies, and an Annexin V-FITC apoptosis kit were purchased from eBioscience (San Diego, CA, USA). The 2′,7′-dichlorodihydrofluorescein diacetate (DCFDA), α-lipoic acid, and 5-methoxytryptamine were purchased from Sigma-Aldrich (St. Louis, MO, USA). The RPMI 1640 medium was purchased from Gibco (Grand Island, NY, USA). The BD Cytofix/Cytoperm buffer was purchased from BD Biosciences (San Diego, CA, USA). Methylcellulose M3534 was purchased from Stem Cell (Vancouer, BC, Canada). Fetal calf serum was purchased from Biological Industries (Kibbutz, Israel). The rabbit anti-γH2AX antibody was obtained from Cell Signaling Technology (Danvers, MA, USA), the rabbit anti-NOX4 antibody from Proteintech (Wuhan, China) and the FITC-conjugated goat anti-rabbit antibodies from Abcam (Cambridge, MA, USA).
The 5-methoxytryptamine-α-lipoic acid (MLA) was synthetized using α-lipoic acid and 5-methoxytryptamine at the drug department of the Institute of Radiation Medicine, CAMS. Melatonin (MLT) was purchased from Tokyo Chemical Industry Co., Ltd. (Shanghai, China). MLA and MLT were dissolved in a 5% carboxymethyl cellulose (CMC) solution.
The 5-methoxytryptamine-α-lipoic acid (MLA) was synthetized using α-lipoic acid and 5-methoxytryptamine at the drug department of the Institute of Radiation Medicine, CAMS. Melatonin (MLT) was purchased from Tokyo Chemical Industry Co., Ltd. (Shanghai, China). MLA and MLT were dissolved in a 5% carboxymethyl cellulose (CMC) solution.
6
Serotonin Receptor Pharmacology Assay
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5HT, E2, 5-methoxytryptamine (MT), and α-methylserotonin maleate salt (MS) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Bovine serum
albumin fraction V was purchased from Merck KGaA (Darmstadt, Germany). Other chemicals of reagent grade were purchased from Wako Pure Chemical Industries
(Osaka, Japan).
albumin fraction V was purchased from Merck KGaA (Darmstadt, Germany). Other chemicals of reagent grade were purchased from Wako Pure Chemical Industries
(Osaka, Japan).
7
Melatonin and its Metabolites Synthesis
8
Drosophila Serotonin Receptor Pharmacology
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The agonists and antagonists used are defined for mammalian preparations and details of binding affinity and efficacy of the various Drosophila 5-HT receptor subtypes are not established. 8-Hydroxy-DPAT hydrobromide (5-HT1A agonist), CP 93129 dihydrochloride (5-HT1B agonist), α-methyl-5-hydroxytryptamine maleate (5-HT2 agonist), and AS19 (5-HT7 agonist) were purchased from Tocris Bioscience (Bristol, UK). 5-Hydroxytryptamine (5-HT), 5-hydroxytryptophan (5-HTP), fluoxetine hydrochloride, para-chlorophenylalanine (PCPA), 5-carboxamidotryptamine maleate salt, 5-methoxytryptamine, BW 723C86 (5-HT2B agonist), R(−)-DOI hydrochloride, (±)-DOI hydrochloride, 1-(3-chlorophenyl)piperazine hydrochloride (m-CPP), and LP-44 (5-HT7 agonist) were purchased from Sigma (St. Louis, MO, USA). All chemicals were dissolved in fly saline. However, AS19 and LP-44 were dissolved in dimethyl sulfoxide (DMSO). Fresh solutions of specific concentrations were made from stock solutions each time before starting an experiment. The DMSO concentration in AS19 was less than 0.01%. The DMSO concentration in LP-44 10 μM, which is the highest concentration employed, was 0.1%.
9
Melatonin Metabolite Quantification in FBS
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