Comparison between YFP-based and APP-labeled axonal lesions with CLARITY at 3 and 24 h after injury. Comparison between YFP-based and APP-labeled axonal.

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From Fig. 1: Photographs of a finished preparation illuminated with transmitted light. The box in i indicates the areas magnified in ii and iii. (ii) shows.

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Comparison between YFP-based and APP-labeled axonal lesions with CLARITY at 3 and 24 h after injury. Comparison between YFP-based and APP-labeled axonal lesions with CLARITY at 3 and 24 h after injury. A, A sagittal view of a cleared brainstem preparation that was also processed for APP IHC (red), visualized here with 2-photon microscopy. For YFP excitation, the 2-photon laser was tuned at 920 nm and for the APP at 780 nm. Scale bar, 300 μm. B, C, Two digital slices from the 3D dataset represent magnifications of boxed areas in A, allowing the visualization of all YFP-filled and APP-labeled axonal lesions at the pyramidal decussation (B) and the crossing of the CST with the RST (C). Panels represent YFP-filled axonal lesions only (green), APP-labeled axonal lesions only (red), or both (red over green). Scale bars: B, 150 μm; C, 200 μm. D, Counts of total numbers of APP-immunoreactive abnormalities at 3 and 24 h after injury (left), densities of APP-positive abnormalities at 3 or 24 h (middle), and rates of colocalization of YFP with APP lesions at the two time points depicted here. Only a subset of YFP-filled axons immunoreact with APP. This is shown by both total counts (compare with counts of Fig. 10E) and percentages shown on the right. Error bars indicate mean ± SEM. Group values were analyzed with one-way ANOVA followed by Tukey's post hoc testing (first three graphs) or Student's t test (fourth graph): *p < 0.05; **p < 0.01; ***p < 0.001. Nikolaos K. Ziogas, and Vassilis E. Koliatsos J. Neurosci. 2018;38:4031-4047 ©2018 by Society for Neuroscience