Dendritic spine density analysis

Dendritic spine density analysis was performed as previously described [47]. Briefly, primary cortical neuron cultures (days in vitro 12–13, DIV 12–13) were transfected with mCAG-IRES-mGFP vector for visualization. The number of dendritic spines was evaluated at DIV 19–20. The fluorescent images were acquired with an LSM 510 confocal microscope (Carl Zeiss, Germany), and the settings were kept consistent for all samples. The dendritic spines were counted on segments of secondary dendrites which are 50–100 μm apart from the center of the cell soma by an investigator blinded to the groups.