Fig. 1.

eCB  signalling regulates cell migration in the RMS.

Sagittal mouse brain slices with GFP-labelled neuroblasts were prepared 5–7 days after in vivo postnatal electroporation of P2 mice with pCX-EGFP, cultured with vehicle or drugs for 2 h and subsequently imaged for 3 h in the same medium. Time-lapse movies made from the descending arm of the RMS in slices treated with different drugs targeting the eCB system (the CB1/2 antagonists AM251 + JTE-907 or the DAGL inhibitor OMDM188, all at 1 μM) were analysed using Volocity. Representative pictures of slices treated with vehicle (A) or the CB1/2 antagonists AM251 + JTE-907 (1 μM each) (B) or OMDM-188 (1 μM) (C) are shown. Arrows follow two neuroblasts in each frame. Insets show magnifications of the neuroblast indicated by the white arrows (A) and (C) or red arrow (B). Representative migratory tracks of 15 cells over 3 h from a control (D), a CB1/2 antagonist-treated (E) or a DAGL inhibitor-treated brain slice (F). White stars mark the tracking end point of each cell. The OB label shows the location of the olfactory bulb in each brain slice. Cells with decreased eCB signalling spent more time immobile (G). Incubation with CB antagonists or DAGLs inhibitors also significantly decreased the percentage of neuroblasts migrating towards the OB (H) and the overall cell displacement (I). Graphs show mean ± s.e.m. (n = 7 brain slices for each condition, ~ 15–30 cells analysed per slice); *p < 0.05, **p < 0.01, ***p < 0.001. Bars, 35 μm for (A–B), 10 μm for insets. (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article.)