Fig. 2

Strong gliosis in 7-month-old BDNF knockout retinas. (A): Staining of retinal cross-sections of 3-month-old mice with an antibody against GFAP (mainly astrocytes, green) and DAPI (cell nuclei, blue). (B): A significantly increased GFAP⁺ area was noted in both knockout groups compared to controls (both: p < 0.050). (C): Gfap mRNA expression was significantly upregulated in the knockout groups (both: p < 0.010). (D): Immunohistological staining of cross-sections of the retina with GFAP (green) and DAPI (cell nuclei, blue). (E): Both knockout groups showed significantly larger GFAP⁺ signal area compared to the control group (both: p < 0.001). (F): Significantly higher Gfap mRNA expression was revealed in CLF and LF retinas (both: p < 0.010). (G): Staining of retinal cross-sections of 3-month-old mice with vimentin (mainly Müller glia, green) and DAPI (cell nuclei, blue). (H): Vimentin staining area remained unaltered. (I): A significant upregulation of Vim mRNA expression was seen in CLF (p < 0.050) but not in LF retinas. (J): Retinal cross-sections were stained with vimentin (green) and DAPI (cell nuclei, blue). (K): Both knockout groups showed a significantly increased vimentin⁺ area compared to the control group (both: p < 0.050). (L): A significant upregulation of Vim mRNA expression was detected in CLF (p < 0.010) as well as in LF mice (p < 0.001). (M): Staining of retinal cross-sections in young mice with nestin (progenitor cells, red) and DAPI (cell nuclei, blue). (N): Area evaluation of nestin showed no differences within all groups. O: Significant upregulation of Nes mRNA expression was noted in the knockout retinas (both: p < 0.050). (P): Cross-sections of older mice are marked with nestin (red) and DAPI (cell nuclei, blue). (Q): Area evaluation of nestin revealed a significant upregulation in the 7-month-old CLF (p < 0.010) and LF animals (p < 0.050) compared to controls. (R): A significant upregulation of Nes mRNA expression was noted in the knockout groups (both: p < 0.010). Values in B, E, H, K, N, and Q are mean ± SEM and each symbol depicts an individual data point. Values in C, F, I, L, O, and R are median ± quartile ± min/max, here the dotted lines represent the relative mRNA expression of the controls. GCL: ganglion cell layer, INL: inner nuclear layer, ONL: outer nuclear layer. *p < 0.050, **p < 0.010, ***p < 0.001. Immunhistological data was analyzed by ANOVA followed by Tukey Honest post-hoc test with n = 6 retinas/group/age, RT-qPCR data was assessed via Pair Wise Fixed Reallocation Randomisation Test© with n = 4–5 samples/group/age. Scale bars = 20 μm.