Rescue of photoreceptors by BDNF gene transfer using in vivo electroporation in the RCS rat of retinitis pigmentosa

　　张萌 莫晓芬 方媛 郭文毅 吴继红 张圣海

　　复旦大学附属眼耳鼻喉科医院 200031

　　Purpose:To investigate the feasibility of introducing brain-derived neurotrophic factor(BDNF)gene into retinal pigment epithelial cells in vivo by electroporation and whether this method can rescue photoreceptors of retinitis pigmentosa in Royal College Surgeons(RCS)rats.

　　Methods：The BDNF-GFP fusion eukaryotic expressing plasmid was constructed and subretinally or intravitreously injected into the eyes of RCS rats followed by in vivo electroporation.The expression of BDNF mRNA and protein was detected by RT-PCR and Western immunoblot analysis.The number of surviving photorecptors was counted and the TdT-dUTP terminal nick-end labeling(TUNEL)method was used to detect the apoptotic retinal cells at different time points after introduction of BDNF plasmid.

　　Results：Treated eyes showed a significantly higher rescue ratio and a lower number of TUNEL-positive photoreceptors than did the control eyes at various time points.

　　Conclusion：These findings provide evidence that electroporation is an effective method for gene transfer into retinal pigment epithelial cells and the rescue of photoreceptors can be achieved by BDNF gene transfection with electroporation.