THC (Sigma Aldrich, St Louis, MO) was obtained under the Federal Drug Enforcement Administration (DEA) license of our collaborator Dr. Bruce Goldberger at the University of Florida. Vehicle control for THC is ethanol (ETOH). Cannabinoid receptor specific agonists, JWH-133, ACEA, and O-1602, were obtained from Tocris (Bristol, UK). Vehicle control for JWH-133 is Tocrisolve (Tocris, Bristol, UK); for ACEA, ETOH; for O-1602, methyl acetate. Cells were exposed to THC using 3 different treatment protocols (Figure 1 ). In treatment 1, monocytes received THC or vehicle control on days -7 and -5 concurrent with MCSF treatment during differentiation into MDM. In treatment 2, MDM were differentiated and treated with THC or vehicle control 30 minutes prior to infection. Finally, in treatment 3, MDM were treated with THC or vehicle control on post infection days 1, 4, and 7. Toxicity of THC or cannabinoid receptor agonists was evaluated by MTS viability assays; briefly, MDM were incubated with CellTiter 96 Aqueous One Solution (Promega, Madison, WI) for 3 hours and absorbance measured at a wavelength of 490 nm using a universal microplate reader (BioTek Instrument Inc, Winooski, VT). Cell viability after THC treatments was 92-105% compared to untreated or vehicle treated cells.
O 1602
Manufactured by Bio-Techne
Sourced in United Kingdom, United States
O-1602 is a research-use only laboratory instrument designed for the measurement and analysis of biological samples. It utilizes a proprietary detection technology to quantify specific analytes or targets within a sample. The core function of O-1602 is to provide accurate and reliable data to support scientific research and exploration.
Automatically generated - may contain errors
Lab products found in correlation
Cid16020046, by Bio-Techne (4 mentions)
Capsazepine, by Merck Group (2 mentions)
Ici 118 551, by Merck Group (2 mentions)
Opti mem, by Thermo Fisher Scientific (2 mentions)
Lipofectamine rnaimax, by Thermo Fisher Scientific (2 mentions)
Nifedipine, by Bio-Techne (1 mentions)
Jwh 015, by Bio-Techne (1 mentions)
Hc 030031, by Bio-Techne (1 mentions)
ω conotoxin mviic, by Bio-Techne (1 mentions)
Pertussis toxin ptx, by Merck Group (1 mentions)
Tocrisolve, by Bio-Techne (1 mentions)
Jwh133, by Bio-Techne (1 mentions)
Celltiter 96 aqueous one solution, by Promega (1 mentions)
O 1918, by Bio-Techne (1 mentions)
Accutase cell detachment solution, by Merck Group (1 mentions)
R methanandamide, by Merck Group (1 mentions)
Sirna targeting, by Santa Cruz Biotechnology (1 mentions)
N acetyl cysteine (nac), by Merck Group (1 mentions)
Lps from salmonella typhimurium, by Merck Group (1 mentions)
Mk 801, by Bio-Techne (1 mentions)
11 protocols using o 1602
1
THC Modulation of HIV Infection
2
Investigating Cannabinoid Receptor Signaling
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The National Institute on Drug Abuse provided Cannabidiol, SR144528, and SR141716. Pertussis toxin (PTX) and capsazepine were purchased from Sigma-Aldrich (St Louis, MO, USA). JWH-015, o-1602, nifedipine, ω-conotoxin MVIIC, and HC-030031 were purchased from Tocris (Ellisville, MO, USA). All compounds were dissolved in dimethyl sulfoxide (DMSO) and added to fresh culture media such that the effective DMSO concentration was 0.1% per well.
3
Investigating Cannabinoid Receptor Signaling
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CBD was bought from Biomol GmbH (Hamburg, Germany). JWH-133 and THC were bought from Bio-Techne GmbH (Wiesbaden, Germany) and Lipomed GmbH (Herne, Germany), respectively. R(+)-methanandamide, capsazepine and NAC were obtained from Sigma-Aldrich (Taufkirchen, Germany). AM251 and AM630 were obtained from Biozol (Eching, Germany). O-1602 and O-1918 were purchased from Tocris Bioscience (Bristol, UK) and Cayman Chemicals (Ann Arbor, Michigan, USA), respectively. SnPPIX was obtained from Enzo Life Sciences (Lörrach, Germany). The transfection reagent Lipofectamine™ RNAiMAX, OptiMEM and hPDGF-BB were purchased from Thermo Fisher Scientific Inc. (Schwerte, Germany). siRNA targeting HO-1 was purchased from Santa Cruz Biotechnology (Heidelberg, Germany; sc-35554). Negative control siRNA was from Qiagen (Hilden, Germany; cat. no. 1022076). Accutase cell detachment solution was obtained from Merck Chemicals GmbH (Darmstadt, Germany).
4
Maternal O-1602 Administration in Mice
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Pregnant C57BL/6
female mice, pregnancy period of 18–19 days, were purchased
from the Air Force Medical University Experimental Animal Center (Xi’an,
China) and housed individually in a temperature- and humidity-controlled
environment, with a 12:12 h light/dark cycle (light on at 07:00 a.m.)
and ad libitum access to food and water. All behavioral tests were
performed during the light period on the designated day of the experiment.
All experimental procedures were approved by the Air Force Medical
University Animal Care and Use Committee (no. 20230702). Efforts were
made to reduce the number of animals used and their suffering. O-1602
(purity >98%) was purchased from Tocris Bioscience (Bristol, UK)
and
dissolved in saline. According to our previous studies,66 (link) the doses of O-1602 (0.1, 0.4, and 2.0 mg/kg,
once a day) were administered via intraperitoneal injection.
female mice, pregnancy period of 18–19 days, were purchased
from the Air Force Medical University Experimental Animal Center (Xi’an,
China) and housed individually in a temperature- and humidity-controlled
environment, with a 12:12 h light/dark cycle (light on at 07:00 a.m.)
and ad libitum access to food and water. All behavioral tests were
performed during the light period on the designated day of the experiment.
All experimental procedures were approved by the Air Force Medical
University Animal Care and Use Committee (no. 20230702). Efforts were
made to reduce the number of animals used and their suffering. O-1602
(purity >98%) was purchased from Tocris Bioscience (Bristol, UK)
and
dissolved in saline. According to our previous studies,66 (link) the doses of O-1602 (0.1, 0.4, and 2.0 mg/kg,
once a day) were administered via intraperitoneal injection.
5
Synthesis and Characterization of Synthetic GPR55 Antagonists
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Synthetic GPR55 antagonists (Fig. 1 ) were synthesized at the Institute for Organic Chemistry Karlsruhe - KIT (Karlsruhe, Germany) [18 (link)] and dissolved in DMSO. The number of the corresponding compounds in the Rempel et al. 2013 paper are: KIT3 is 14, KIT17 is 37, and KIT21 is 41. ML193 and O-1602 were obtained from Tocris Biosciences. LPS from Salmonella typhimurium (Sigma Aldrich, Deissenhofen, Germany) was resuspended in sterile phosphate-buffered saline (PBS, 5 mg/mL) as stock and subsequently used at a final concentration (10 ng/mL) in the cultures.![]()
Molecular structure of the synthesized compounds
6
Investigating Immune Cell Signaling Pathways
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O-1602 and CID16020046 were purchased from Tocris Bioscience (Ellisville, MO, USA). R-phycoerythrin (PE)-conjugated mouse anti-human Mac-1 (clone ICRF44; BD) antibody (Cat No. 555388) and PE-conjugated mouse IgG isotype control (clone MOPC-21) antibody (Cat No. 555749) were obtained from BD Biosciences (San Diego, CA, USA). Various signal pathway inhibitors were purchased from EMD Serono (Rockland, MA, USA) and Sigma-Aldrich (St. Louis, MO, USA).
7
Electrophysiological Experiments Drug Regimen
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Drugs used for the electrophysiological experiments were applied by dissolving them to the desired final concentration in the bathing ACSF. The concentrations of the various drugs were as follows: AM281 (2 μM), 5′-Iodoresiniferatoxin (I-RTX, 1 μM), CNQX (10 μM), MK-801 (25 μM), orlistat (5 μM), N-palmitoylethanolamine (PEA, 10 μM), O-1602 (10 μM), CID-16020046 (CID-1602, 20 μM), (S)-3,5-DHPG (50 μM) (from Tocris, Bristol, UK). Bicuculline (10 μM) (from Sigma-RBI, St. Louis, USA). Bicuculline, CNQX, (S)-3,5-DHPG and MK-801 were dissolved in water. AM281, I-RTX, orlistat, O-1602 and CID-16020046 were dissolved in DMSO. PEA was dissolved in ethanol. The intraneuronal delivery of orlistat was performed dissolving the drug (orlistat, 5 µM) in the internal solution.
8
Synthesis of Enantiomeric Adrenergic Compounds
9
Preparation and Use of GPR55 Agonists and Antagonists
10
Synthesis and Use of Fenoterol Analogs
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(R,R′)-Fenoterol [(R,R′)-Fen], (R,S′)-4′-methoxy-1-naphthylfenoterol [(R,S′)-MNF], and (R,R′)-MNF were synthesized as described previously28 (link). The following compounds from Tocris Bioscience (Ellisville, MO, USA) were used for cell treatments: O-1602 (GPR55 agonist), Tocrifluor T1117 (T1117, fluorescent GPR55 agonist), myristoylated protein kinase inhibitor-(14–22)-amide (PKI), and forskolin (adenylyl cyclase activator). Salmeterol (selective β2-AR agonist) and ICI-118,551 (ICI, selective β2-AR inverse agonist) were purchased from Sigma-Aldrich (St-Louis, MO, USA).
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