Transcriptome changes in Olig2-3 over-expression retina ganglion cells (RGCs) by AAV-Olig1/2.
To gain mechanistic insights into the effects of Olig family, we performed gene expression profiling analysis in RGCs. Transcriptome changes in Olig1-3 over-expression RGCs showed up-regulating genes related to axon projection development and regeneration-associated genes (RAGs), such as Stmn2, Gal, Gap43, and Sprr1a. Overall design: Adult male mice were anesthetized with avertin (300-500mg/kg) and positioned in a stereotaxic apparatus. Under microscopic illumination, injections were made through the sclera and retina with a 5µL Hamilton syringe with a 32-gauge needle, avoiding injuring lens for excluding the false-positive possibility of regeneration. In this work, all pacaged AAV2 virus (10^13 gc/mL, 1.5µL) were injected into the vitreous chamber to infect RGCs. 14 days after ONC, we separated RGCs, which were transfected with AAV-Olig2-3 or AAV-GFP, by magnetic beads coupled with CD90.2 antibody and picked 15~20 cells by Unipick in each sample.