Experimental scheme of retroviral injection paradigm. Adult mice were stereotactically co‐injected with the CAG‐dnLEF‐IRES-GFP (dnLEF) and CAG‐RFP (control) MMLVs and were sacrificed 17 and 42 dpi.
Representative images of the DG with transduced neurons at 17 and 42 dpi. CAG‐dnLEF‐IRES-GFP transduced cells in green, and CAG‐RFP transduced cells in red. Scale bar = 100 μm.
Quantification of RFP‐positive and GFP‐positive cells show reduced survival of neurons transduced with dnLEF (P = 0.0286; n = 4 animals).
Morphology analysis of surviving neurons shows no difference in dendritic length (P = 0.1004) and number of branch points (P = 0.8654) between dnLEF and control neurons, but a reduction in complexity (Sholl analysis; P = 0.0176; control: n = 18 cells from four animals, dnLEF: n = 15 cells from four animals).
Schematic representation of the retroviral paradigm used to analyze survival of canonical Wnt signaling gain of function in neural progenitors.
Representative images of the dentate gyrus of control and β‐catex3 mice with GFP expressing transduced neurons (green) and the immature marker doublecortin (DCX) (gray) at 42 dpi. Scale bar = 100 μm.
Quantification of GFP‐positive cells in control and β‐catex3 animals show reduced survival of neurons with stable expression of β‐cat from the stage of fast‐proliferating progenitor stage on (P = 0.0286; n = 4 animals).
Morphology analysis of surviving GFP‐positive neurons shows no difference in dendritic length (P = 0.0503) and number of branch points (P = 0.5051), but differences in complexity in Sholl analysis (P = 0.0240) between genotypes (control: n = 20 cells from four animals, dnLEF: n = 15 cells from four animals).
Data information: Data represented as mean ± SEM, significance was determined using two‐way ANOVA for Sholl analysis and two‐tailed Mann–Whitney
‐test for all other analyses, and significance levels were displayed in GP style (*
< 0.0332).
