Jwh133

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.

Cannabidiol (CBD) was generously provided by the NIDA Drug Supply Program and was dissolved in the 5% cremophor. Sucrose (Sigma-Aldrich) was dissolved in 0.9% physiological saline to achieve 5% concentration that was delivered in a volume of 0.1 or 0.02 ml onto a food trough. JWH133, AM251, and AM630 were purchased from Tocris Bioscience. JWH133 was dissolved in soya oil-based Tocrisolve-100 (Tocris Bioscience, USA). AM251 and AM630 were dissolved in 5% cremophor and were administered intraperitoneally (i.p.).

Intrathecal injection was performed in lightly restrained unanesthetized mice as previously described [11 (link)]. We injected 5 μl of drug [JWH-133 (Tocris, UK), AM-630 (Tocris, UK), or WIN55,212-2 (Cayman Chemical, Ann Arbor, MI) and/or vehicle [ethanol: alkamuls EL-620 (Rhodia, Cranbury, NJ): saline in a volume ratio of 1:1:8]. The data of Figure 1 includes animals that were injected twice (Days 24 and 28 after the first MOG injection) using a balanced crossover design. Animals receiving vehicle on day 24 received drug on day 28, and vice versa. Group means of either vehicle or drug on day 24 and day 28 did not differ, and so were combined for final analysis. The data of Figure 2 were obtained on day 28 after two consecutive i.t. injections, separated by 15 min: first AM-630 or vehicle and then JWH-133, followed by behavioral testing sessions.