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Fig. 8
Figure 8. Retinal spatiotemporal expression pattern of Mmp-9 protein after ONC in adult zebrafish. A: Representative picture showing presence of active Mmp-9 in zebrafish retinal extracts, as confirmed by labeling of both pro (92 kDa) and active (76 kDa) Mmp-9 in zebrafish embryo lysates (left). A,B: Western blotting for Mmp-9 on retinal extracts at different timepoints postinjury indicates a peak in active Mmp-9 expression at 14 dpi compared to sham-operated controls. Data represent at least three retinal samples per timepoint and are shown as mean ± SEM (*P < 0.05). C,D: Immunostainings for Mmp-9 on retinal sections show an increased Mmp-9 signal in three prominent IPL strata at 14 dpi. E-H: Double staining with SV2 confirms expression of Mmp-9 in a subset of IPL synapses. F-H: High-magnification pictures of double (F) and single (G,H) stainings. For all immunostainings, DAPI (blue) was used as a nuclear counterstain. ONC, optic nerve crush; dpi, days postinjury; NFL, nerve fiber layer; RGCL, retinal ganglion cell layer; IPL, inner plexiform layer; INL, inner nuclear layer; OPL, outer plexiform layer; ONL, outer nuclear layer; PRL, photoreceptor layer. Scale bars = 20 µm in C-E; 5 µm in F-H.