Rescue of axotomized retinal ganglion cells by BDNF gene electroporation in adult rats.
OBJECTIVE
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.
Mo
Xiaofen
X
Department of Ophthalmology and Visual Science, Graduate School of Medicine, Chiba University, Inohana 1-8-1, Chuo-ku, Chiba 260-8670, Japan. xfmo@hotmail.com
Yokoyama
Akiko
A
Oshitari
Toshiyuki
T
Negishi
Hisanari
H
Dezawa
Mari
M
Mizota
Atsushi
A
Adachi-Usami
Emiko
E
D000818:Animals; D017209:Apoptosis; D019771:Axotomy; D015153:Blotting, Western; D019208:Brain-Derived Neurotrophic Factor / Q000235:genetics* / Q000378:metabolism; D002452:Cell Count; D002470:Cell Survival / Q000502:physiology*; D018274:Electroporation / Q000379:methods*; D019084:Fluorescent Antibody Technique, Indirect; D015870:Gene Expression; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D020287:In Situ Nick-End Labeling; D008297:Male; D009900:Optic Nerve / Q000601:surgery; D051381:Rats; D017208:Rats, Wistar; D012165:Retinal Ganglion Cells / Q000166:cytology* / Q000502:physiology; D014162:Transfection
D019208:Brain-Derived Neurotrophic Factor