Rescue of Photoreceptors by BDNF Gene Transfer Using in vivo Electroporation in the RCS Rat of Retinitis Pigmentosa

Abstract

Purpose: : To investigate the feasibility of transfecting brain-derived neurotrophic factor (BDNF) gene into retinal pigment epithelium (RPE) cells and retinal ganglion cells (RGCs) in vivo by electroporation for preventing photoreceptor degeneration in the Royal College Surgeons (RCS) rats of retinitis pigmentosa.

Methods: : The BDNF-GFP fusion expression plasmid was constructed, and then transferred into the RPE cells or RGCs of RCS rats by subretinal or intravitreal injection routes followed by in vivo electroporation. The expression of BNDF mRNA and protein were detected by reverse transcription polymerase chain reaction technique (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 apoptosis of the retinal cells at different time points after introduction of BDNF plasmid.

Results: : After the electrointroduction, treated eyes showed a significantly higher number of rescued photoreceptors,and a lower number of TUNEL-positive photorecptors than the control ones at various time points, especially in the group of both RPE cells and RGCs were transfected with BDNF plasmid.

Conclusions: : These findings provide evidence that electroporation is an effective method for transferring genes into RPE cells and RGCs and rescue of photorecptros in RCS rats can be achieved by BDNF gene transfection with in vivo electroporation.

Keywords: gene transfer/gene therapy • retinal pigment epithelium • retinal degenerations: cell biology