AM6545, a peripherally-restricted CB1R neutral antagonist, was given by IP injection at 10 mg per kg (Northeastern University Center for Drug Discovery, Boston, MA, USA). Devazepide (Tocris, Bristol, UK), a CCKA receptor antagonist, was given IP at 0.3 mg per kg. Both drugs were dissolved in vehicle consisting of 7.5% DMSO, 7.5% Tween80, and 85% sterile saline, and warmed in a water bath to ensure solubility. All control conditions were matched, using vehicle in place of drugs and injections occurred 1 h prior to behavior recording (1,600 h). A 72-h washout period was allowed between drug treatments. JZL184 (Tocris, Bristol, UK), a potent inhibitor of monoacylglycerol lipase (MGL), was used to prevent monoacylglycerol hydrolysis in the diacylglycerol lipase (DGL) assay and to validate our MGL assay (described below). Tetrahydrolipstatin (Tocris, Bristol, UK), a lipase inhibitor used routinely to study DGL activity (Gregg et al., 2012 (link); Jung et al., 2012 (link)), was used to validate our DGL assay.
Devazepide
Manufactured by Bio-Techne
Sourced in United Kingdom
Devazepide is a selective cholecystokinin (CCK) type B receptor antagonist that is used in scientific research. It is a white crystalline powder. Devazepide has a molecular formula of C17H15ClN2O2 and a molecular weight of 330.77 g/mol.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (10 mentions)
Fura 2 am, by AAT Bioquest (3 mentions)
Dulbecco s modified eagle s medium dmem f 12 medium, by Thermo Fisher Scientific (2 mentions)
Pkillerredpm vector, by Evrogen (2 mentions)
Ampicillin, by CWBIO (2 mentions)
Kanamycin, by CWBIO (2 mentions)
Jetprime transfection reagent, by Polyplus Transfection (2 mentions)
Dexamethasone (dex), by Merck Group (2 mentions)
Jzl184, by Bio-Techne (1 mentions)
Tetrahydrolipstatin, by Bio-Techne (1 mentions)
Aminoguanidine ag, by Merck Group (1 mentions)
Sq22536, by Merck Group (1 mentions)
Forskolin, by Merck Group (1 mentions)
Pyrrolidine dithiocarbamate pdtc, by Merck Group (1 mentions)
Ym022, by Bio-Techne (1 mentions)
Coelenterazine, by Promega (1 mentions)
Mitotracker red fm, by Thermo Fisher Scientific (1 mentions)
Lyso tracker red, by Beyotime (1 mentions)
Clozapine n oxide, by Bio-Techne (1 mentions)
Cck 8, by Bio-Techne (1 mentions)
8 protocols using devazepide
1
Pharmacological Validation of Lipase Assays
2
Inflammatory Signaling Pathway Modulators
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Sulfated cholecystokinin octapeptide (CCK-8), lipopolysaccharide (LPS; Escherichia coli serotype O111:B4), aminoguanidine (AG), dexamethasone (DEX), pyrrolidine dithiocarbamate (PDTC), SQ-22,536, H-89, and forskolin were obtained from Sigma-Aldrich (St. Louis, MO, USA). The CCK-Rs antagonists, devazepide, and YM-022 were purchased from Tocris Bioscience (Ellisville, MO, USA).
3
Optimized Bioluminescence Imaging Protocol
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Sulphated cholecystokinin octapeptide (CCK) and CCK1R antagonist devazepide were from Tocris Cookson (Bristol, UK). Dulbecco’s Modified Eagle’s Medium (DMEM) / F12 medium was bought from InVitrogen (Shanghai, China). Goods buffer 4-(2-hydroxyethyl)-1-piperazine-ethane-sulfonic acid (HEPES) was from Calbiochem (Darmstadt, Germany). Fura-2 AM was from AAT Bioquest (Sunnyvale, CA, USA). JetPRIME transfection reagent was from PolyPlus-transfection (New York, USA). Sulphonated aluminium phthalocyanine (SALPC) was from Frontier Scientific Inc. (AlPcS-834, Logan, UT, USA). Cell-Tak and Agar (BactoTM) were from BD Biosciences (Bedford, MA, USA). Fetal bovine serum (FBS) was from Thermo Scientific (Shanghai, China). pKillerRedPM vector was bought from Evrogen (Moscow, Russia). pNL1.1.CMV (NanoLuc / CMV) vector containing the NanoLuc luciferase gene under the control of the cytomegalovirus (CMV) promoter and coelenterazine were from Promega (Madison, WI, USA). PEF1A-IRES-Neo vector was from Addgene (Watertown, MA, USA). Ampicillin and Kanamycin were from CWBio (Beijing, China). Endotoxin-free plasmid extraction kit and DH5à competent cells were from TianGen Biochemicals (Beijing, China).
4
Subcellular Localization of CCK Signaling
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Sulfated cholecystokinin octapeptide (CCK) and CCK1R antagonist devazepide were from Tocris Cookson (Bristol, UK). Dulbecco’s modified Eagle’s medium (DMEM)/F12 medium was bought from Invitrogen (Shanghai, China). Fura-2 AM was from AAT Bioquest (Sunnyvale, CA, USA). JetPRIME transfection reagent was from PolyPlus-transfection SA (New York, NY, USA). Fetal bovine serum (FBS) was from Thermo Scientific (Shanghai, China). pKillerRedPM vector was bought from Evrogen (Moscow, Russia). Ampicillin and kanamycin were from CWBio (Beijing, China). Endotoxin-free plasmid extraction kit and DH5à competent cells were from TianGen Biochemicals (Beijing, China). MitoTracker™ Red FM was from Invitrogen (Carlsbad, CA, USA). LysoTracker Red was from Beyotime (Shanghai, China).
5
Intraperitoneally Administered Drugs for In Vivo Studies
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Drugs for in vivo use were prepared in sterile saline and administered intraperitoneally (IP). Clozapine-N-oxide (CNO; Tocris Bioscience, Bristol, UK; Cat. No. 4936) was administered at 0.3 mg/kg. Devazepide (Tocris Bioscience, Cat. No. 2304) was dissolved in DMSO, further diluted with sterile saline and administered at 1 mg/kg, IP, 40 min before CNO injection or optogenetic photostimulation. CCK-8 (Tocris, Cat. No. 1166) and SR 27,897 (CCK-A antagonist; Tocris Bioscience, Cat. No. 27897) were dissolved in artificial cerebrospinal fluid (aCSF).
6
Devazepide-Induced Cell Apoptosis Assay
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Human BC 5637 cells were provided by Shanghai Cell Bank of the Chinese Academy of Sciences, Shanghai, China. Devazepide (obtained from Tocris Bioscience, UK) was dissolved in absolute ethanol to 50 mM. Dulbecco’s modified Eagle’s medium (DMEM), trypsin, ethylenediaminetetraacetic-acid solution, and fetal bovine serum (FBS) were obtained from GIBCO, USA. An annexin V, fluorescein isothiocyanate (FITC) apoptosis detection kit and a cell counting kit-8 (CCK-8) were purchased from Dojindo (Kumamoto, Japan). CyclinD1 antibody (BS1741), Bcl-2-associated X protein (Bax) antibody (BS2538), goat anti-rabbit immunoglobulin G (IgG) (H + L)-FITC and horseradish peroxidase (HRP) labeled anti-rabbit secondary antibody were purchased from Bioworld, USA. Ki67 antibody (ab16667), Tubulin antibody (ab7291), PARP1 antibody (ab191217), and Cleaved Caspase-3 antibody (ab32042) were acquired from Abcam, UK.
7
Zebrafish Model of Intestinal Inflammation
8
Imaging of CCK1R Receptor Dynamics
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Sulfated cholecystokinin octopeptide (CCK), CCK1R antagonist devazepide were from Tocris Cookson (Bristol, United Kingdom). MEM amino acid mixture (50×), DMEM/F12, penicillin/streptomycin, Opti-MEM, and MitoTracker® Green FM were from InVitrogen (Shanghai, China). 4-(2-Hydroxyethyl)-1-piperazine-ethane-sulfonic acid (HEPES) was from Calbiochem (Darmstadt, Germany). Fura-2 AM was from AAT Bioquest (Sunnyvale, CA, United States). Transfection reagent X-tremeGENE HP was from Roche (Mannheim, Germany). Cell-Tak and Agar (BactoTM) were from BD Biosciences (Bedford, MA, United States). Fetal bovine serum (FBS) was from Thermo Scientific (Shanghai, China). pKillerRedmem and pKillerReddMito were bought from Evrogen (Moscow, Russia). Yeast extract and tryptone were from MERCK (Darmstadt, Germany). Endotoxin-free plasmid extraction kit and DH5à competent cells were from TianGen Biochemicals (Beijing, China). Restriction enzymes (BamHI, EcoRI, and XhoI) were from Takara (Beijing, China). Goat and rabbit anti-CCK1R polyclonal antibody, 2nd ab (FITC- or DyLight 488-labeled) were from Abcam (Cambridge, United Kingdom). Hoechst 33342 was from DojinDo (Beijing, China). Photosensitizer SALPC was from Frontier Scientific (AlPcS-834, Logan, UT, United States).
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