Transcriptional profiling of adult retinal ganglion cells during optic nerve regeneration

Objective: To identify genes that are differentially expressed in retinal ganglion cells undergoing axon regeneration after optic nerve injury. Adult mice that express cyan-fluorescent protein (CFP) in RGCs were treated with pro-regenerative treatment after optic nerve injury. The treated RGCs were selected by FACS (CFP+mcherry) and their RNA profiles were analyzed. Adult CFP mice received AAV2-shPten-mcherry or AAV2-mcherry virus 2 weeks before optic nerve crush. Immediately after nerve injury, AAV2-shPten-injected mice received recombinant oncomodulin plus CPT-cAMP (T) and AAV2-mcherry-injected mice received saline intraocularly (C). RGCs were selected by FACS 1, 3, 5 days (D1, D3, D5) after nerve crush and treatment. Normal RGCs (Nor.) received AAV2-mcherry only. To find distinctive regulation after pro-regenerative treatment, samples from the treatment group were compared with that of same time point control group: D1.C vs. D1.T; D3.C vs. D3.T; D5.C vs. D5.T. All of the nerve-injured samples were also compared with normal RGCs.