A Novel Constitutively Active , p.(R33P) Variant in RAB11A Associated with Intellectual Disability Promotes Neuritogenesis and Affects Oligodendroglial Arborization

Figure 4

The expressions of RABs in oligodendrocytes and effect of RAB11A-R33P expression on oligodendrocyte morphology. (a) RAB11A, 5, and 7 are expressed in cultured mouse oligodendrocytes. Western blotting of cultured oligodendrocyte lysates was performed, and signal bands confirming the expressions of RAB11A, 5, and 7 are shown. ß-Actin was used as a loading control. (b) Representative images of primary cultured oligodendrocytes expressing RAB11A WT, -R33P, -S25N, or -Q70L variants. At 96 h, after the induction of differentiation, cells were stained with antibodies against GFP and MBP. The top: representative images of oligodendrocytes expressing EGFP-tagged RAB11A WT and variants. The middle: representative images of oligodendrocytes stained with MBP merged with EGFP images. MBP images display arborized cytoplasmic processes in each oligodendrocyte. The bottom: binary images of MBP staining visualizing detailed arborization. The insets show high-power magnification views of boxed areas. Scale bars: 50 μm (the top and middle), 10 μm (the bottom inset). (c, d) Graphs presenting Sholl analysis data on oligodendrocytes expressing the various RAB11A. The mean of the total numbers of intersections in each oligodendrocyte (c), and the mean of the radial lengths of the outermost rings intersecting the oligodendroglial process (d) are shown ( in each group). Oligodendrocytes infected with an empty EGFP vector were used as control. ; versus RAB11A WT.

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