Figure 5.

Cul3 homozygous deletion and heterozygous reduction lead to unchanged excitatory synapses in primary cultured hippocampal neurons. (A) Representative images of excitatory synaptic puncta in WT, heterozygous, and homozygous fCul3 cultured hippocampal neurons transduced with either AAV-GFP (WT-GFP, Het-GFP, Hom-GFP) or AAV-Cre-GFP (WT-Cre, Het-Cre, Hom-Cre). Scale bars represent 50 μm for the larger images and 20 μm for the magnified dendrite images. (B) Presynaptic excitatory synapse marker vGluT1 puncta density is comparable in both heterozygous and homozygous Cul3 knockout neurons (Het-Cre and Hom-Cre) compared to control heterozygous and homozygous fCul3 neurons transduced with AAV-GFP (Het-GFP and Hom-GFP). (C) Postsynaptic excitatory synapse marker PSD95 puncta density is unchanged in both heterozygous and homozygous Cul3 knockout neurons (Het-Cre and Hom-Cre) compared to control heterozygous and homozygous fCul3 neurons transduced with AAV-GFP. (D) Percentage of co-localized PSD95/vGluT1 puncta is unchanged in homozygous Cul3 knockout neurons (Hom-Cre) compared to homozygous fCul3 neurons transduced with AAV-GFP (Hom-GFP). *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001, ordinary two-way AVONA with Dunnett’s multiple comparisons test; graphs depict mean ± SEM. N = 45 neurons from 3 separately derived neuronal cultures (15 neurons for each group).