Dendritic spine density analysis

To study the quantity and morphology of dendritic spines of pyramidal cells in the CA1 region of the hippocampus, brains were removed from the animals and incubated in a series of solutions using the FD Rapid GolgiStain kit (FD Neurotechnologies), according to the manufacturer’s instructions. Following cryoprotection of the whole brains with Solution C (see kit), tissue was sectioned coronally at 200 μm thickness and then viewed under a light microscope. Dendritic arbors of each cell were analyzed as previously described [29]. The spine density was calculated as the number of spines (defined as protrusions of the dendritic membrane regardless of shape) along a 30-μm dendritic terminal segment. A total of 10 different neurons were analyzed for each animal per group.