Figure 5

Impact of the pharmacological blockade of A_2ARs in status epilepticus-induced cell proliferation and migration of adult-born neurons. (a) Six/seven weeks old rats were injected either with saline (Control group) or with pilocarpine (Status Epilepticus group), 30 min after the administration of scopolamine methylbromide (2 mg/kg, i.p.). Only animals reaching at least stage 4 seizures were considered. Convulsions were suppressed with diazepam (10 mg/kg, i.p.) 2 h after the occurrence of the first stage 4 seizure. The rats from each group were assigned into two subgroups: one subgroup was daily injected with the selective antagonist of A_2AR, SCH58261 (0.1 mg/kg i.p.), and the other with vehicle (0.2% Tween-20 in saline), starting 10 h after the onset of status epilepticus. All the animals were i.p. injected with BrdU at 24 h (200 mg/kg), 7 days (200 mg/kg) and 8 days (100 mg/kg) after status epilepticus. BrdU⁺-cells in the granule cell layer (GCL) per dentate gyrus (DG) was evaluated at 42 days post-status epilepticus by (b) immunohistochemical analysis of BrdU (green), NeuN (magenta) and GFAP (cyan). Scale bar, 300 μm. (c) Status epilepticus induced a significant increase in BrdU⁺-cells in the GCL per DG in vehicle-injected rats. SCH58261 significantly modified status epilepticus-induced increase in BrdU⁺-cells density (*P < 0.05 status epilepticus x SCH58261 interaction; two-way ANOVA). Post-hoc analysis revealed that status epilepticus did not significantly increase BrdU⁺-cells density in rats administered with SCH58261, but also there was no significant difference between status epilepticus-vehicle and status epilepticus-SCH58261. (d) Vehicle-injected rats that experienced status epilepticus displayed a higher relative percentage of BrdU⁺-cells in the outer two-thirds of the GCL in comparison with the Control group. No interaction was found between SCH58261 and status epilepticus factors (two-way ANOVA). The data are mean ± SEM of the number of the BrdU⁺-cells in GCL or the relative percentage of cells in the inner (1/3) and in the outer (2/3) parts of GCL. Control-Vehicle, n = 7; Control-SCH58261, n = 4; Status Epilepticus-Vehicle, n = 12; Status Epilepticus-SCH58261, n = 6. ML—molecular layer; GCL—granule cell layer; SGZ—subgranular zone. *P < 0.05, **P < 0.01 and ***P < 0.001, two-way ANOVA with Sidak’s test.