Fig 2: Immunocytochemical analyses of DAGL and MAGL in primary neuronal (left column), microglial (middle column) and astrocyte (left columns) cell cultures.In neurons, DAGL was distributed in the perikarya and in neuronal processes. VGLUT1 was co-localized with DAGL suggesting a close surveillance of glutamate by DAGL (arrows). MAGL was distributed in primary neuronal cultures but displayed no overlap with VGLUT1 (arrow). Microglial cells were labeled with DAGL in their cell bodies after LPS treatment. DAGL expression was stronger than in controls (arrow). whereas, MAGL expression in microglia was clearly visualized under control condition around the nucleus and at the outer edges (arrows) of the cells the labeling disappeared under LPS treatment (arrow). Astrocytes were not found immunoreactive for DAGL in fibrillary astrocytes under control condition, however DAGL was weakly induced after LPS treatment. Protoplasmic astrocytes displayed a robust DAGL immunoreaction in controls and after LPS stimulation as visualized in droplets throughout the cell body (arrows). MAGL was induced in both fibrillary and protoplasmic astrocytes after LPS treatment (arrows). Bar = 20 µm.

Fig 3: Representative immunoblots (A) and relative protein levels of components of the eCB signaling system [CB1 (B), NAPE-PLD (C), FAAH (D), DAGLα (E), DAGLβ (F), MAGL (G), NAPE-PLD (H), DAGLα/MAGL (I), and DAGLβ/MAGL (J)] in the mouse cerebellum after chronic pretreatment (vehicle and cocaine) and acute treatment (vehicle and cocaine). Histograms represent the mean ± SEM (n = 8/group). Two-way ANOVA: ns, no significance; *P < 0.05, **P < 0.01. Bonferroni: *P < 0.05, **P < 0.01 vs. vehicle–vehicle group.

Fig 4: Fatty acid amide hydrolase (FAAH) and monoacylglycerol lipase (MAGL) protein levels in adult rats from the adequate (control) and the ISR condition. FAAH protein levels (A) were decreased specifically in the amygdala of ISR rats but were unchanged in the mPFC, striatum and thalamus. MAGL protein levels (B) were significantly reduced in the striatum of ISR rats compared to control rats but were unchanged in the mPFC, amygdala and thalamus. Data are indicated as means ± SEM (control/ISR: n = 10 each; *P ≤ 0.05).

Fig 5: Enzymatic regulation of 2-AG after PPT.(A) Dagl and Magl mRNA regulation after PPT in the entorhinal cortex and the hippocampus after PPT. Dagl was increased after 24 hpl of PPT in the DG. (n = 3, *p <0.05) (B) DAGL (open cycles) and MAGL (open cycles) protein regulation after PPT in the entorhinal cortex and the hippocampus after PPT. DAGL was decreased after 24 hpl of PPT in the DG compared to the respective control (closed cycles). (n = 3, *p<0.05).