Figure 3.

Effect of WWL70 and JZL184 on [³H]–2-AG hydrolysis and 2-AG accumulation in intact neurons in primary culture. (a) [³H]–2-AG hydrolysis by intact primary neurons after 30 min pretreatment with WWL70 (10 μM) or JZL184 (1 μM). The data are expressed as % of control hydrolysis (pretreatment with 0.1% DMSO). (b,c) Levels of 2-AG in intact primary neurons pretreated for 30 min with WWL70 (10 μM), JZL184 (1 μM) or vehicle (0.1% DMSO, control; b) and stimulated with glutamate (100 μM) and carbachol (1 mM) for 2.5 min, after which lipids were extracted and 2-AG amounts measured by GC-MS (c). Treatment with glutamate plus carbachol led to a 2.5-fold increase in 2-AG amounts (in fmol per 100,000 cells: 18 to 44). Data are shown as mean ± s.e.m. of 3–6 independent experiments. Experiments were performed in triplicate for the hydrolysis assay and in duplicate for 2-AG quantification by GC-MS, using cells from independent cell culture preparations. *P < 0.05, **P < 0.01, ***P < 0.001 (ANOVA one-way, Bonferroni's post test).