Neurogenesis assay

The neurogenic effect of chronic fluoxetine treatment in combination with D1 receptor agonist/antagonist was assessed as described [22]. Briefly, the mice were labeled with BrdU solution (200 mg/kg) for 2 h prior to sacrifice by perfusion. Preparation of brain sections and immunostaining were done as described above, and stereological quantitation of BrdU-positive cells was performed as described [22]. Briefly, serial coronal sections (40 µm) were cut through the entire hippocampus in its rostrocaudal extension on a cryostat. Every sixth section throughout the hippocampus was processed for BrdU immunohistochemistry. An experimenter blinded to experimental conditions counted all BrdU-labeled cells in the granule cell layer and the SGZ of the dentate gyrus in the total 12 sections from the individual mouse. The total number of BrdU-labeled cells per section was determined and multiplied by 6 to obtain the total number of cells per dentate gyrus.