Drugs were prepared as 1000× stocks and stored at −20°C. To induce homeostatic scaling, neurons were treated with 20 μm bicuculline methobromide (BCC; Tocris, 20 mm stock in water) or 1 μm tetrodotoxin citrate (TTX; Abcam, 1 mm stock in water) for 48 h (Fig. 1 ). In some live-cell imaging experiments, 20 μm BCC or 1 μm TTX was applied acutely for 10–60 min. For Figure 2 , neurons were treated with 20 μm BCC for 4, 12, 24, or 48 h to monitor the process of homeostatic down-scaling. For Figure 3 , neurons were treated for 48 h with a combination of 1 μm MTEP hydrochloride (Tocris, 1 mm stock in DMSO) and 100 nm JNJ 16259685 (Tocris, 100 μm stock in DMSO) to block mGluR1/5 signaling. For Figure 4 , neurons were treated for 48 h with 100 nm PF3845 (Cayman, 100 μm stock in DMSO) or 2.5 μm PF3845 (Cayman, 2.5 mm stock in DMSO) to inhibit FAAH and to elevate AEA and related NAEs. For Figure 5 , neurons were treated for 6 h with 50 nm AM251 (Cayman, 50 μm stock in DMSO), or for 10–60 min with 500 nm AM251 (Cayman, 500 μm stock in DMSO) to block CB1 signaling. For Extended Data Figure 5-1 , neurons were treated for 10–60 min with 50 μm D-AP5 (Cayman, 50 mm stock in water) to block NMDA signaling.
Pf3845
Manufactured by Cayman Chemical
Sourced in United States
PF3845 is a laboratory product that functions as a chemical compound. Its core purpose is to serve as a research tool for scientific investigations. Detailed specifications and intended applications are not available at this time.
Automatically generated - may contain errors
Lab products found in correlation
Gw6471, by Cayman Chemical (2 mentions)
Anandamide aea, by Cayman Chemical (2 mentions)
Urb937, by Cayman Chemical (2 mentions)
Am251, by Cayman Chemical (2 mentions)
Lipofectamine rnaimax, by Thermo Fisher Scientific (2 mentions)
Fura 2 am, by Thermo Fisher Scientific (2 mentions)
Arachidonic acid, by Cayman Chemical (1 mentions)
Gw9662, by Cayman Chemical (1 mentions)
Urb597, by Cayman Chemical (1 mentions)
Sr141716, by Cayman Chemical (1 mentions)
Sr144528, by Cayman Chemical (1 mentions)
O 1918, by Cayman Chemical (1 mentions)
Fp tamra, by Thermo Fisher Scientific (1 mentions)
Lipopolysaccharide, by Merck Group (1 mentions)
Arachidonic acid, by Merck Group (1 mentions)
D ap5, by Cayman Chemical (1 mentions)
Jnj16259685, by Bio-Techne (1 mentions)
Mtep hydrochloride, by Bio-Techne (1 mentions)
Oea d2, by Cayman Chemical (1 mentions)
D4 aea, by R&D Systems (1 mentions)
7 protocols using pf3845
1
Homeostatic Scaling of Neuronal Activity
2
Serine Hydrolase Activity-Based Profiling
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The serine hydrolase activity-based protein profiling probe (ABPP probe), FP-TAMRA, was purchased from Thermo Fisher Scientific (Waltham, MA, USA). Small interfering RNAs (FlexiTube SI02736706 for fatty acid amide hydrolase (FAAH) gene knockdown and Allstars Negative Control) were purchased from Qiagen (Tokyo, Japan), and transfection reagent Lipofectamine RNAiMAX was purchased from Thermo Fisher Scientific. All inhibitors, antagonists, and reagents, including PF3845, URB597, SR141716, SR144528, GW9662, GW6471, O1918, anandamide (AEA), and arachidonic acid (AA), were purchased from Cayman Chemical (Ann Arbor, MI, USA).
Radioactive anandamide [ethanolamide-1,2-14C] was purchased from American Radiolabeled Chemicals, Inc (Saint Louis, MO, USA). Other chemicals including lipopolysaccharide were purchased from Sigma-Aldrich (St. Louis, MO, USA).
Radioactive anandamide [ethanolamide-1,2-14C] was purchased from American Radiolabeled Chemicals, Inc (Saint Louis, MO, USA). Other chemicals including lipopolysaccharide were purchased from Sigma-Aldrich (St. Louis, MO, USA).
3
Acute THC Effects on c-Fos and Locomotion
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For acute THC-induced c-Fos and locomotion experiments, THC (25 mg/ml; Sigma-Aldrich) was dissolved in a vehicle solution composed of 5% Tween 80 and 95% saline to a final concentration of 0.5 mg/ml. The diluted THC was injected by intraperitoneal route at 10 ml/kg for a final dosage of 5 mg/kg. For CPP experiments, THC was dissolved in a vehicle solution composed of 5% Tween 80 and 95% saline to a final concentration of 0.1 mg/ml. The diluted THC was injected by intraperitoneal route at 10 ml/kg for a final dosage of 1 mg/kg. The FAAH inhibitor PF-3845 (Cayman Chemical) was resuspended in dimethyl sulfoxide (DMSO) to a concentration of 10 mg/ml. The concentrated PF-3845 was diluted 1:1:8 (PF-3845/Tween 80/saline) to a final concentration of 1 mg/ml. The diluted PF-3845 was injected by intraperitoneal route 2 hours before each THC injection at 10 ml/kg for a final dosage of 10 mg/kg.
4
Synthesis and Characterization of Endocannabinoids
5
Pharmacological Compounds for Endocannabinoid Research
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GW6471, AM251, URB937, PF3845, AMC-AEA, LEI-401, PEA, d4-PEA, OEA, d2-OEA, 2-AG, and d5-2-AG were purchased from Cayman Chemical (Ann Arbor, MA). NGF was from Gibco (Waltham. MA). AEA and d4-AEA were from R&D Systems (Minneapolis, MN). Capsaicin was from Sigma-Aldrich (St. Louis, MO), Fura-2AM and PED-A1 were from Thermo Fisher (Waltham, MA) and Cyclosporin A was from TCI (Portland, OR).
6
Foot-Shock Stress and Cannabinoid Modulation
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All studies were carried out in accordance with the National Institute of Health Guide for the Care and Use of Laboratory Animals and approved by the Vanderbilt University Institutional Animal Care and Use Committee. Male ICR mice (4–7 weeks old) were housed on a 12:12 light–dark cycle with lights on at 0600 (Harlan, Indianapolis, IN, USA). All experiments were conducted during the light phase. Food and water were available ad libitum.
Foot-shock stress occurred 24 h before behavioral testing and consisted of six 0.7 mA foot-shocks delivered 1 min apart using a MED Associates fear-conditioning chamber (St. Albans, VT, USA). Each 2-s shock was preceded by a 30-s auditory tone. Drugs were administered 2 h before behavioral testing. The FAAH inhibitor PF-3845 (10 mg kg−1, Cayman Chemical, Ann Arbor, MI, USA) and the CB1 receptor antagonist rimonabant (3 mg kg−1, NIMH Drug Supply Program) were dissolved in dimethylsulfoxide (Sigma Aldrich, Milwaukee, WI, USA) and intraperitoneally injected at a volume of 1 μl g−1.
Foot-shock stress occurred 24 h before behavioral testing and consisted of six 0.7 mA foot-shocks delivered 1 min apart using a MED Associates fear-conditioning chamber (St. Albans, VT, USA). Each 2-s shock was preceded by a 30-s auditory tone. Drugs were administered 2 h before behavioral testing. The FAAH inhibitor PF-3845 (10 mg kg−1, Cayman Chemical, Ann Arbor, MI, USA) and the CB1 receptor antagonist rimonabant (3 mg kg−1, NIMH Drug Supply Program) were dissolved in dimethylsulfoxide (Sigma Aldrich, Milwaukee, WI, USA) and intraperitoneally injected at a volume of 1 μl g−1.
7
THC Conditioned Place Preference with FAAH Inhibitor
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To test the effect of FAAH inhibition on THC CPP, adolescent female C57BL/6J mice (WT) were treated with either vehicle (1:1:8, DMSO/Tween 80/saline) or the FAAH inhibitor PF-3845 (10 mg/kg, ip; Cayman Chemical), 2 hours before each THC injection during THC CPP training sessions. Mice received no PF-3845 treatment on test day.
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