Figure 11.

The C2B domain present in both RIM3γ and RIM4γ is sufficient to rescue the knock-down phenotype. A, Hippocampal neurons were transduced at DIV 1 with viral particles expressing GFP (Control) or shRIM3 together with either RIM3γ (nonresistant) or RIM4γ. Neurons transduced at DIV 1 with viral particles expressing GFP (Control) or shRIM4 in combination with either RIM4γ (nonresistant) or RIM3γ. B, C, Quantification shows that the phenotype caused by knock-down of either RIM3γ (B) or RIM4γ (C) can be rescued by overexpression of the respective other isoform. (A, right column) Neurons were transduced with viral particles expressing shRIM3 or shRIM4 in combination with the respective RIM3/4γ-C2B domain. D, The quantification revealed that the C2B domain is sufficient to restore dendritic complexity almost to control levels. All neurons were analyzed at DIV 14 using confocal microscopy. Sholl analysis was performed to quantify the loss of neuronal processes. One-way ANOVA; ***p < 0.001.