Figure 6

Effects of CB1 absence on neuronal maturation. By using animals at different points of time after BrdU injection a detailed time course of neuronal maturation has been established. At 24 h after BrdU the number of BrdU-positive cells was increased in the mutant mice but at 4 weeks the number of BrdU-positive cells was reduced compared to the controls. The percentage of NeuN-positive cells was also reduced resulting in a net reduction of adult neurogenesis in the mutants. When we looked at the 24 h time point we found a relative reduction in the number of proliferative DCX-positive cells At an intermediate 7d time point, the increased proliferation in CB1^-/- animals had yielded to a strong reduction in BrdU positive cells compared to controls (A). To investigate early stages of neuronal maturation, we used Nestin-GFP-reporter mice and injected the CB1 antagonist AM251. At 1 h after BrdU application counts of BrdU-positive cells reflect S-phase entry. AM251-treatment increased the number of BrdU-positive cells compared to vehicle controls. At 24 h the numbers had roughly doubled, reflecting a completed cell cycle. Phenotypic analysis revealed that this increase was largely accounted by type-2b cells and later DCX-positive cells, whereas type-2a was even reduced. At 48 h control values for BrdU began to be higher than in the AM251-treated mice, leading to an almost two-fold reduction in AM251-treated mice at 7d (B); * p ≤ 0.05.