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. 2020 Feb 27;13:29. doi: 10.3389/fnmol.2020.00029

Figure 2.

Figure 2

Overexpression of p110δ impairs dendritic complexity. (A) Representative tracings of primary cultured rat hippocampal neurons co-transfected with Venus-GFP and p110δ or control expression plasmids on DIV7 and immunostained for GFP and myc (not shown) on DIV10. Colored overlays in control images are representative examples of primary (green), secondary (yellow), and tertiary (blue) dendritic branches. (B) Results of Sholl analysis comparing neurons from control (n = 18) and p110δ-OE (n = 26) cultures. (C,D) Quantification of control (n = 19) and p110δ-OE neurons (n = 28) treatment effects on the mean number and length of primary, secondary, and tertiary dendritic branches. Length measurements were compared by taking an overall average dendrite length for each subtype/treatment [primary branches, n = 146 (control), 201 (p110δ-OE); secondary branches, n = 294 (control), 255 (p110δ-OE); tertiary branches, n = 146 (control), 105 (p110δ-OE)]. (E) Representative images of primary cultured rat hippocampal neurons transfected with p110δ or control expression plasmid on DIV7 and immunostained for myc-tag (red) and p110δ (green) on DIV10. (F) Quantification of average p110δ immunofluorescent intensity [n = 17 (control), 22 (p110δ-OE) neurons]. (G) Representative images of primary cultured rat hippocampal neurons transfected with p110δ or control expression plasmid on DIV7 and immunostained for myc-tag (red) and phosphorylated Akt (p-Akt Ser473; gray) on DIV10. Imaging conditions to detect p-Akt Ser473 under control conditions resulted in overexposure of signal in neurons transfected with p110δ-OE vector. OE, overexpression. *P-value of <0.05, **P-value of <0.01, and ***P-value <0.001. Data are represented as means ± SEM.