Fig. 7
From: Optic nerve as a source of activated retinal microglia post-injury
Naive optic nerve of the CX3CR1YFP-creER:CD11cGFP mouse was well-populated with microglia and generated a strong response to injury. a GFPhi microglia were numerous in naïve optic nerve (A), and GFPlo microglia densely populated the same naïve optic nerve as shown in (B). The Z-stack of flat mounted optic nerve shown in panels A and B was obtained by scanning through the optic nerve to find its center, and then stacking six 3 μm optical sections centered in the middle of the nerve. b The optic nerve at 31 days post-ONC remained densely populated. CD11cGFP reporter = green; DAPI = blue; CX3CR1YFP reporter = yellow. c Quantitation of retinal myeloid cells by flow cytometry of retina and optic nerve from naïve and day 10 post-ONC donors. An optic nerve crush led to increases in GFPhi and GFPlo microglia in retina and optic nerve. Cells were gated on viable CD45medCD11bhiLy6G− cells with doublet exclusion. GFPhi and GFPlo cell counts were limited to CX3CR1-YFPhi cells. Mean ± SD. NS, not significant; *, p < 0.05; **, p < 0.01; ***, p < 0.001. Statistical analysis by one-way ANOVA with Tukey HSD post-test. 6–11 mice/group
