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. 2008 Jul 16;28(29):7387–7398. doi: 10.1523/JNEUROSCI.1942-08.2008

Figure 5.

Figure 5.

Early SCLIP inhibition blocks PC dendritic development at 7 and 14 div. A, Classification of PC differentiation stages in cerebellar organotypic cultures. Rat cerebellar slices were immunostained for CaBP after 3 d (stage 1), 7 d (stage 2), and 14 d (stages 3 and 4) in culture. PCs first display a fusiform bipolar morphology referred to as stage 1, and then exhibit short perisomatic processes characteristic of stellate stage 2. They finally display several perisomatic dendritic processes and eventually a single typical dendritic tree, respectively defined as stages 3 and 4. Scale bar, 20 μm. B, Morphologies of PCs at 7 div after early infection with Lv-shCtrl or Lv-shSCLIP. Cerebellar organotypic cultures were infected just after plating and immunostained for GFP (green) and CaBP (red) 7 d later. Whereas PCs transduced with Lv-shCtrl (visualized by GFP) are in stage 2 and display short perisomatic protrusions as do untransduced PCs (red), PCs transduced with Lv-shSCLIP exhibit a round morphology, without any perisomatic processes but the axon (arrowheads). Scale bar, 20 μm. C, Quantified results of four combined independent experiments illustrating the effect of SCLIP inhibition on PC dendritic differentiation stages at 7 div. Whereas almost all PCs transduced with Lv-shCtrl are in stage 2, >90% of PCs transduced with Lv-shSCLIP are in a round stage. Shown are mean values ± SEM. D, Morphologies of PCs at 14 div after early infection with Lv-shCtrl or Lv-shSCLIP. Cerebellar organotypic cultures were infected just after plating and immunostained for GFP (green) and CaBP (red) 14 d later. In contrast to untransduced PCs or PCs transduced with Lv-shCtrl, which exhibit a large and branched dendritic arbor, PCs transduced with Lv-shSCLIP still display a round morphology (asterisks) or exhibit very short fingered processes (arrows) emerging from the cell body. Note that their axons are, however, not affected by SCLIP inhibition (arrowheads). Scale bar, 20 μm. E, Quantified results of four combined independent experiments illustrating the effects of SCLIP inhibition on PC dendritic differentiation stages at 14 div. Whereas most of PCs transduced with Lv-shCtrl are in stages 3 and 4, ∼75 and 25% of PCs transduced with Lv-shSCLIP are in a round or “digitated” stage, respectively. Values shown are mean ± SEM.