Rimonabant hydrochloride (SR 141716A) and anandamide (in TocrisolveTM 100, 1:4 ratio of soya oil/water and emulsified with the block co-polymer Pluronic F68) were purchased from Tocris Bioscience (Bristol, United Kingdom). TocrisolveTM 100 was used as vehicle in the anandamide studies (Tocris Bioscience, Bristol, United Kingdom). Rimonabant was dissolved in a vehicle of Tween 80 (5% volume/volume, v/v), DMSO (20% v/v), and sterile saline (75% v/v). The volume percentages refer to the final rimonabant solution. Both rimonabant and anandamide were administered intraperitoneally (ip) at a volume of 1 ml/kg. Cannabis cigarettes (5.7% Δ9-THC, 0.02% cannabidiol) were kindly provided by the NIDA Drug Supply Program. The cannabis cigarettes were stored at -20°C. About 24 h before being used, the cigarettes were removed from the freezer and placed in an airtight humidity chamber with a small dish (10 x 10 cm) containing 0.5 cm of water (95% humidity) and kept at room temperature (22–24°C). The cigarettes were used within one hour of being removed from the humidity chamber.
Tocrisolve 100
Manufactured by Bio-Techne
Sourced in United Kingdom, United States
Tocrisolve 100 is a water-soluble formulation of the lipophilic compound Tocrisolve. It is designed to improve the solubility and bioavailability of lipophilic molecules for in vitro and in vivo applications.
Automatically generated - may contain errors
Lab products found in correlation
Pluronic f68, by Bio-Techne (1 mentions)
A769662, by Bio-Techne (1 mentions)
T ampk, by Cell Signaling Technology (1 mentions)
2 deoxy d glucose 2 dg, by Merck Group (1 mentions)
Jzl184, by Bio-Techne (1 mentions)
Urb597, by Merck Group (1 mentions)
Am251, by Cayman Chemical (1 mentions)
Dimethyl sulfoxide (dmso), by Cayman Chemical (1 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)
Tween 80, by Merck Group (1 mentions)
Gw7647, by Bio-Techne (1 mentions)
Indomethacin, by Merck Group (1 mentions)
Lps from escherichia coli o111 b4, by Merck Group (1 mentions)
Pertussis toxin ptx, by Merck Group (1 mentions)
Am251, by Bio-Techne (1 mentions)
Jwh133, by Bio-Techne (1 mentions)
Am630, by Bio-Techne (1 mentions)
Sucrose, by Merck Group (1 mentions)
9 protocols using tocrisolve 100
1
Cannabinoid Compound Pharmacology Protocol
2
Apoptosis Pathway Biomarkers Analysis
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Antibodies (Abs) against apoptosis inducing factor (AIF), phosphorylated AMPK (pAMPK), and total AMPK (tAMPK) were purchased from Cell Signaling (USA). Abs against endonuclease G (endo G) and α-tubulin were purchased from Abcam (UK). The anti-nitrotyrosine Ab was purchased from Upstate Biotechnology (USA). A769662 and Tocrisolve 100 were purchased from Tocris Bioscience (Bristol, UK). The 2-deoxy-D-glucose (2DG) was from Sigma-Aldrich (USA). All other reagents were purchased from Calbiochem (USA).
3
Cannabinoid Receptor Agonists and Inhibitors
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CB1 receptor agonist WIN 55212-2 (WIN, Tocris, 0.1 and 0.5 mg/kg body weight for the intraperitoneal injection, 1 and 2 μg/μl for micro-infusion into the vHip) (Martellotta et al., 1998 (link); Shen et al., 2019 (link)) was dissolved in 50% DMSO (Sigma-Aldrich) and 50% normal saline solution. Arachidonylcyclopropylamide (ACPA, Abcam, 5 or 10 mg/kg body weight for the intraperitoneal injection, 4 ng/μl for micro-infusion into the vHip) (Jafari-Sabet and Karimi, 2017 (link); Shafaroodi et al., 2004 (link); Srisai et al., 2017 (link)) was dissolved in 100% Tocrisolve 100 (Tocris). The FAAH inhibitor URB597 (5 ng/μl, Sigma-Aldrich) (Haller et al., 2009 (link)) was dissolved in 5% DMSO and 95% normal saline solution. MAGL inhibitor JZL184 (2 μg/μl, Tocris) (Folkes et al., 2020 (link)) was dissolved in 1:1 DMSO and Tween 80 (Sigma-Aldrich) and diluted by the normal saline solution to 2% DMSO and 2% Tween 80. The CB1 receptor antagonist AM251 (2.5 ng/μl, Cayman) (Shen et al., 2019 (link)) was dissolved in 25% DMSO and 75% normal saline solution. Drug doses were based on the literature and on pilot experiments. Solutions were freshly prepared and were administrated at a volume of 0.1 ml for the intraperitoneal injection and 0.8 μl per side for the infusion into the vHip and dorsal striatum.
4
Synthesis and Administration of OEA and CC7
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OEA [N-(cis-9-octadecenoyl)-ethanolamine] and GW7647 [2-(4-(2-(1-cyclohexanebutyl)-3-cyclohexylureido)ethyl)phenylthio)-2-methylpropionic acid] were purchased from Tocris Bioscience (Bristol, UK). The disubstituted sulfamides CC7 (N-octadecyl-N′-propylsulfamide) and CC12 (N-2-adamantyl-N′-propylsulfamide) were synthesized as previously described [17] (link).
For in vitro molecular biology studies, the drugs were dissolved in dimethyl sulfoxide (DMSO). For in vivo studies in rodents, the drugs were dissolved in 20% Tocrisolve™-100 (Tocris Bioscience, Bristol, UK) and 80% physiological saline. OEA and CC7 were administered intraperitoneally (i.p.) at doses of 5 mg/kg in a volume of 1 mL/kg body weight in rats and 10 mL/kg body weight in mice. A solution of 0.6% acetic acid was prepared in physiological saline.
For in vitro molecular biology studies, the drugs were dissolved in dimethyl sulfoxide (DMSO). For in vivo studies in rodents, the drugs were dissolved in 20% Tocrisolve™-100 (Tocris Bioscience, Bristol, UK) and 80% physiological saline. OEA and CC7 were administered intraperitoneally (i.p.) at doses of 5 mg/kg in a volume of 1 mL/kg body weight in rats and 10 mL/kg body weight in mice. A solution of 0.6% acetic acid was prepared in physiological saline.
5
Inflammatory Mediator Protocols
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LPS from Escherichia coli O111 :B4 (dissolved in isotonic NaCl; Sigma-Aldrich), indomethacin (suspended in 1 % Tween 20; Sigma-Aldrich), PGE2 (dissolved in Tocrisolve 100 and diluted with isotonic NaCl; Sigma-Aldrich), and arachidonic acid (dissolved in Tocrisolve 100, Tocris) and their respective vehicles were administered in a volume of 1 ml/kg.
6
Neuronal Pharmacological Treatments and Protocols
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TC, MLA, TEA and pertussis toxin (PTX) were purchased from Sigma-Aldrich. AA, Chelerythrine chloride (Ch Cl-), 3-(2,4-Dichlorophenyl)-4-(1-methyl-1H-indol-3-yl)-1H-pyrrole-2,5-dione (SB 216763) and Tocrisolve 100 were purchased from Tocris. 5,8,11,14-Eicosatetraynoic acid (ETYA) was purchased from Santa Cruz Biotech. Drugs were dissolved in dimethyl sulfoxide (DMSO) or DM according to solubility, and then added to culture media shortly after neurons were plated (~ 1 h). The next day, three washes with DM followed by replacement with fresh DM or CM was performed to remove drugs prior to experiments (~ 1 h). 0.1% DMSO and Tocrisolve 100 vehicle controls were performed for all experiments, as appropriate.
7
Cannabidiol Delivery and Receptor Antagonists
8
Topical CB2R Agonists and Inverse Agonist
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CB2R agonists, RO6871304 [33 (link)], RO6871085 [30 ], and the CB2R inverse agonist, RO6851228 [32 (link)], were provided by Hoffman-la Roche Ltd. (Basel, Switzerland), and were dissolved for topical administration in Tocrisolve™ 100 (Tocris Bioscience, Ellisville, MO, USA), which is a 1:4 ratio of soya oil:water, that is emulsified with the block co-polymer Pluronic F68. RO6871304, RO6871085, or RO6851228 were applied topically on the left eye (1.5% w/v; 5 μL/animal)) directly after injection of LPS in the same eye. Control animals received vehicle (Tocrisolve) (5 μL/animal) only. HU910 was dissolved for i.v. administration in mixed micelles.
9
Investigating NP-1's Anorectic Properties
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NP-1 was generously provided by Noscira (formerly Neuropharma, Tres Cantos, Spain). NP-1 was dissolved in a vehicle of 10% Tocrisolve-100 (Tocris Bioscience, Bristol, UK) and 90% physiologic saline and administered intraperitoneally (i.p.) at a dose of 5 mg/Kg, chosen based on its previously established anorectic properties10 (link). Drugs were injected at a volume of 1 ml/kg.
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