Figure 7

Overexpression of normal or mutant CREST affects complexity of dendritic tree in primary hippocampal neurons. (A) CREST-GFP is largely confined in the nucleus when expressed in primary mouse neurons but is also found in multiple dot-like aggregates in the processes in some cells. Untagged CREST displays nuclear distribution in neurons. CREST-GFP or untagged CREST was co-transfected into primary hippocampal neurons together with dsRed2 to visualise neuronal morphology and allowed to express for 48 hours prior to analysis. (B) Total dendritic length in micrometers measured in mouse hippocampal neurons expressing CREST variants. Mouse primary hippocampal neurons were co-transfected with vectors to express GFP (to visualize neuronal morphology) and each of the untagged CREST variants and allowed to express the proteins for 48 hours prior to analysis. Control neurons were transfected with GFP-expressing vector only. The bar charts show means ± s.e.m. for at least 100 neurons per variant from four independent experiments (*p < 0.05, ***p < 0.001). (C) Sholl analysis of the same neurons as in B. Number of dendrite intersections of 15-μm spaced shells as a function of the radial distance from the soma was plotted. **p < 0.01, ***p < 0.001. Scale bar, 50 μm.