Figure 4

RGC count in retina flatmounts and retina crosssections. (A) Representative Nissl-stained flatmounts of CO and EAE retinas reveal a loss of neurons (yellow arrows) in EAE retinas. The white arrows show glia cells and the arrowheads endothelial cells. Scale bar: 20 μm.(B) Quantification of neurons on retinal flatmounts (n = 8 eyes/group). The number of neurons (P< 0.0001) was significantly reduced in the EAE group after 23 days. (C) Immunohistochemical staining of CO and EAE retinas with Brn-3a (RGCs, red) and DAPI (nuclei, blue) 23 days post immunization. The merged picture shows a reduction of Brn-3a⁺ stained RGCs (arrows) in the GCL of EAE animals. (D) Quantification of Brn-3a⁺ cells in the retinas of CO and EAE animals (n = 5 to 7 eyes/group). RGC numbers were significantly reduced in the retinas of EAE mice (CO: 32.5 ± 1.5 cells/mm, EAE: 26.7 ± 1.9 cells/mm, P = 0.0170). Scale bar: 25 μm. *: p<0.05; ***: p<0.001. CO, control; EAE, experimental autoimmune encephalomyelitis; GCL, ganglion cell layer; INL, inner nuclear layer; IPL, inner plexiform layer; RGC, retinal ganglion cell.