Fig. 5.

Mouse neurons and microglial cells produce four endocannabinoids. a–d, Mouse neurons were incubated with ionomycin (iono; 5 μm) for 2.5 and 5 min or with glutamate (Glut; 100 μm) and carbachol (Carb; 1 mm) for 2.5 min, lipids were extracted from chloroform from cells plus adjacent media and purified by HPLC; AEA (a), 2-AG (2AG; b), HEA (c), and DEA (d) were then quantified by CI-GC/MS.e–h, BV-2 cells were incubated with ionomycin (5 μm) for increasing periods of time, and endocannabinoids were quantified. prot, Protein. The arrowindicates that EGTA (5 mm) prevents the ionomycin response.i–l, Mouse microglial cells were incubated with ionomycin (5 μm) for 2.5 and 5 min or with ATP (1 mm) for 10 min, and endocannabinoids were quantified. Values are means ± SEM of 6–32 independent endocannabinoid quantifications, each performed on one 100 mm dish of cells. *p < 0.05; **p < 0.01; significantly different from basal amount (see Table 1) (ANOVA followed by Dunnett's post hoc test).