Figure 6. Endogenous Dynamin-3 is Required for Postsynaptic Positioning of the Endocytic Zone.
(A) Dynamin-3 knock-down causes loss of the EZ. Hippocampal neurons expressing either dynamin-3 shRNA (d3RNAi-2) or a scrambled shRNA control (scramble) together with clathrin-DsRed were fixed and stained for VGLUT1. Scale bar, 5 µm.
(B) High magnification views of glutamatergic synapses (VGLUT1, green) and postsynaptic clathrin (clathrin-DsRed, red). Postsynaptic neurons expressed the indicated shRNA constructs. Scale bar, 1 µm.
(C) Quantification of endocytic zone (EZ) negative synapses in neurons expressing dynamin-3 shRNAs (d3RNAi-2, d3RNAi-4), dynamin-2 shRNA (d2RNAi-2) or a scrambled RNAi control. Co-expression of RNAi-resistant dynamin-3 cDNAs (Dyn3-2*, Dyn3–4*, Dyn3–4*-P800L, Dyn3-ΔGED) is indicated. Data are means ± SEM. n = 36, 41, 40, 13, 18, 25, 11 and 16, respectively; ** p < 0.001 relative to scrambled shRNA control, t-test.