Rescue of axotomized retinal ganglion cells by BDNF gene electroporation in adult rats

Xiaofen Mo, Akiko Yokoyama, Toshiyuki Oshitari, Hisanari Negishi, Mari Dezawa, Atsushi Mizota, Emiko Adachi-Usami

Research output: Contribution to journalArticlepeer-review

73 Citations (Scopus)

Abstract

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′-tetramethylin-docarbocyamine percholorate (dil) 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.

Original languageEnglish
Pages (from-to)2401-2405
Number of pages5
JournalInvestigative Ophthalmology and Visual Science
Volume43
Issue number7
Publication statusPublished - 2002 Jul 9
Externally publishedYes

ASJC Scopus subject areas

  • Ophthalmology
  • Sensory Systems
  • Cellular and Molecular Neuroscience

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