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Figure 5 | Molecular Brain

Figure 5

From: Automated 4D analysis of dendritic spine morphology: applications to stimulus-induced spine remodeling and pharmacological rescue in a disease model

Figure 5

A PI3 kinase inhibitor rescues spine morphology in neurons from Fmr1 knockout mice. (a) Hippocampal neurons cultured from wild type (WT) or Fmr1 knockout (KO) mice were treated with vehicle or a PI3 kinase inhibitor (LY294, 10 μM) for 72 hours starting at 15 DIV. Neurons were transfected with a plasmid encoding Lifeact-ruby at 17 DIV and fixed 24 hours later. The images depict representative dendritic regions from deconvolved z-series images (scale bar is 5 μm). (b) Spine density was measured using our automated approach for WT and KO neurons treated with either vehicle or LY294 (n = 55-60 neurons; ANOVA [F = 3.996, P = 0.009]; post-hoc Fisher's LSD: *P = 0.017, #P = 0.028). Cumulative distributions of (c) spine head width, (d) length, and (e) volume were plotted for each group (Kolmogorov-Smirnov test: head width [WT vs. KO: P = 0.002, WT vs. WT LY294: P = 0.235, KO vs. KO LY294: P = 0.009], spine length [WT vs. KO: P = 0.009, WT vs. WT LY294: P = 0.537, KO vs. KO LY294: P = 0.014], spine volume [WT vs. KO: P < 0.001, WT vs. WT LY294: P = 0.158, KO vs. KO LY294: P < 0.001]). (f) Spines were classified as stubby, mushroom, and thin based on the automated geometric measurement, and the values were plotted as percentage of total spines per treatment group (n = 55 - 60 neurons; ANOVA with post-hoc Fisher's LSD: *P = 0.043, #P = 0.006, ‡P = 0.043).

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