Figure 5.

EphrinB3 mRNA and protein expression in the adult retina and on RGC axons. A, EphrinB3 mRNA expression in the ganglion cell layer and the inner nuclear layer of the retina from a normal adult mouse. [See D for location of ganglion cell layer (GCL) and inner nuclear layer (INL).] B, EphrinB3 mRNA expression in the ganglion cell layer and the inner nuclear layer 4 d after optic nerve injury. C, EphrinB3 mRNA expression in the ganglion cell layer and the inner nuclear layer 12 d after optic nerve injury. D, Retinal section from a normal adult mouse exposed to the EphrinB3 sense control probe. E, EphrinB3 mRNA expression in the optic nerve of a normal adult mouse. A faint hybridization signal is detected in cells distributed in a row-like manner. F, EphrinB3 mRNA expression in the optic nerve of an adult mouse 4 d after optic nerve injury. G, EphrinB3 mRNA expression in the optic nerve of an adult mouse 12 d after optic nerve injury. H, Anti-EphrinB3 immunoblot. A faint signal is detected in the protein sample from uninjured optic nerves (UI). A band at ∼65 kDa, the expected size of EphrinB3, is present in the optic nerve samples 8 d after injury (8). I, Uninjured optic nerve section after staining using anti-EphrinB3 antibody. There is diffuse staining particularly at the ONH region. J, Optic nerve (ON) tissue section 12 d after injury stained with anti-EphrinB3 antibody. A strong signal was detected in the optic nerve proximal to the injury site. CT, Connective tissue; L, lesion site. K, Section from the same optic nerve shown in J after staining with a secondary antibody alone. L, Recombinant EphB3-Fc protein injected into the adult optic nerve binds to RGC axons. EphB3-Fc was detected using an anti-Fc antibody. M, Fc control protein injected into the adult optic nerve is distributed as diffuse punctate aggregates and does not result in apparent RGC axon binding. Scale bars: A–G, I–K, 100 μm; L, M, 20 μm.