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. 2020 Apr 27;21(9):3079. doi: 10.3390/ijms21093079

Figure 5.

Figure 5

Fingolimod-phosphate (FTY720-P) shows mild rescue in neuronal morphology of young cortical neurons from Cdkl5-/y mice. Young DIV7 cortical cultures from mice carrying a Cdkl5-/y mutation or littermate wild type (WT) controls, were grown in presence of DMSO alone or 10 nM FTY720-P in the growth medium. (A) The diagram shows the scheme of treatment from culture preparation on DIV0, treatment from DIV1 to fixation on DIV7. (B) Micrographs display representative DIV7 MAP2 positive cortical neurons. The WT and Cdkl5-/y neurons were both treated either with DMSO or FTY720-P. Scale bar: 50 μm. (C) Neurite complexity for both genotypes treated with DMSO and FTY720-P was analyzed and plotted as number of neurite intersections against the distance from the soma. F value for comparison between the four sets is described in the graph. The Sholl curve for following genotype and treatment sets are also displayed separately (D) WT (black) v/s Cdkl5-/y (dark red), (E) Cdkl5-/y neurons treated with 10nM FTY720-P (rosa) v/s DMSO controls (dark red), (F) WT neurons treated with 10nM FTY720-P (gray) v/s DMSO controls (black). (G) Total intersections, (H) number of neurites and (I) total length of neurites as computed for WT neurons treated with DMSO (black) or FTY720-P (gray) and Cdkl5-/y neurons treated with DMSO (dark red) or FTY720-P (rosa). Data in graphs is represented as mean + SEM. Numbers in the bars show total number of neurons analyzed for each genotype and treatment, obtained from ≥3 sets of independent experiments. Two-way ANOVA followed by Bonferroni post-hoc test was used in (A). For (G,H) and (I) one-way ANOVA with Bonferroni post-hoc was used. Denotations for significance are * p < 0.05, **** p < 0.0001.