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(Investigative Ophthalmology and Visual Science. 2002;43:2401-2405.)
© 2002 by The Association for Research in Vision and Ophthalmology, Inc.

Rescue of Axotomized Retinal Ganglion Cells by BDNF Gene Electroporation in Adult Rats

Xiaofen Mo1, Akiko Yokoyama1, Toshiyuki Oshitari1, Hisanari Negishi1, Mari Dezawa2, Atsushi Mizota1 and Emiko Adachi-Usami1

1 From the Department of Ophthalmology and Visual Science, Graduate School of Medicine, Chiba University, Chiba, Japan; the 2 Department of Anatomy, Yokohama City University School of Medicine, Yokohama, Japan.

PURPOSE. To determine whether the brain-derived neurotrophic factor (BDNF) gene can be transfected into retinal ganglion cells (RGCs) by electroporation and whether axotomized RGCs can be rescued after transfection by BDNF in adult rats.

METHODS. Mouse BDNF cDNA was injected intravitreally followed by in vivo electroporation in adult rats. The expression of BDNF in RGCs was confirmed by Western immunoblot analysis and immunohistochemistry. After introduction of BDNF cDNA, the survival of axotomized RGCs was estimated by the TdT-dUTP terminal nick-end labeling (TUNEL) method and measured by counting the number of RGCs that were labeled retrogradely by 1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyamine percholorate (diI) applied to the superior colliculus (SC).

RESULTS. Eyes with injection of the BDNF gene followed by in vivo electroporation showed a significantly higher level of expression of BDNF in the RGC layer, a higher rescue ratio, and a lower number of TUNEL-positive cells than the control samples.

CONCLUSIONS. These findings demonstrate that electroporation is an effective method for the direct delivery of genes into RGCs, and that the BDNF gene transferred into RGCs by in vivo electroporation can protect axotomized RGCs against apoptosis.




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