Figure 4.

OTUD7A Localizes to Dendritic Spines and Regulates Spine Density

(A) OTUD7A localizes to dendritic spines. Mouse primary cortical cultures were co-transfected with pEGFP-c1 and 3xFLAG-OTUD7A on DIV 7. On DIV 14, immunofluorescent imaging was performed, with GFP in green (Alexa488) and FLAG in red (Cy3). The bottom right corner of each image shows a magnification of the inset box.

(B) Primary cortical neurons from Otud7a-null mice show decreased total spine density compared to wild-type neurons, which are rescued by overexpression of Otud7a. To compare between genotypes, primary cortical neurons from Otud7a-null mice or wild-type littermates were transfected with pEGFP-c1 and stained with GFP antibody. As a rescue experiment, primary neurons from Otud7a-null mice were transfected with 700 ng 3xFLAG-OTUD7A per 24-well. We investigated 42 neurons/dendrites from 9 null mice, 41 neurons/dendrites from 3 wild-type mice, and 31 neurons/dendrites from 9 null mice in the rescue experiments. Top: representative examples of dendritic segments of the immune-stained neurons. Bottom: quantification of dendritic spines. Statistical significance was assessed using two-tailed t test. Data are presented as mean ± SEM (^∗p < 0.05, ^∗∗p < 0.01, ^∗∗∗p < 0.001).