Abstract |
We developed an in vivo electroporation method to introduce foreign genes into retinal ganglion cells (RGCs). After the intravitreous injection of the plasmid gene (20 mug), five electric pulses (6 V/cm, 100 ms duration) were each delivered twice with 5 min interval to the rat eye using a contact lens-type electrode (cathodal) attached to the cornea and a needle electrode (anodal) inserted to the middle of the forehead. The efficiency of the genetic introduction into RGCs and tissue damage to the eyeball was evaluated using a green fluorescent protein (GFP) gene, TUNEL and histological observation. DiI retrograde labeling revealed that 24.4 +/- 4.7% of all RGCs were electrointroduced with the GFP gene. TUNEL and histological analysis showed a few tissue damages in the cornea, lens and retina. To confirm whether this method can actually rescue damaged RGCs, glial cell line-derived neurotrophic factor ( GDNF) was electrointroduced into RGCs after optic nerve transection. After the electrointroduction, a significant increase in the number of surviving RGCs was observed 2 and 4 weeks after the optic nerve transection, and the decrease of caspase 3 and 9 was detected by RT-PCR. These results suggest that this method may be useful for the delivery of genes into RGCs with simplicity and minimal tissue damage.
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Authors | H Ishikawa, M Takano, N Matsumoto, H Sawada, C Ide, O Mimura, M Dezawa |
Journal | Gene therapy
(Gene Ther)
Vol. 12
Issue 4
Pg. 289-98
(Feb 2005)
ISSN: 0969-7128 [Print] England |
PMID | 15616607
(Publication Type: Journal Article)
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Chemical References |
- Gdnf protein, rat
- Glial Cell Line-Derived Neurotrophic Factor
- Nerve Growth Factors
- Green Fluorescent Proteins
- Casp3 protein, rat
- Casp9 protein, rat
- Caspase 3
- Caspase 9
- Caspases
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Topics |
- Animals
- Axotomy
- Caspase 3
- Caspase 9
- Caspases
(genetics)
- Cell Survival
- Electrodes
- Electroporation
(instrumentation, methods)
- Gene Expression
- Genetic Therapy
(methods)
- Glial Cell Line-Derived Neurotrophic Factor
- Green Fluorescent Proteins
(genetics)
- Injections
- Male
- Nerve Growth Factors
(genetics)
- Optic Nerve
(surgery)
- Rats
- Rats, Wistar
- Retinal Ganglion Cells
(metabolism, pathology)
- Reverse Transcriptase Polymerase Chain Reaction
- Transgenes
- Vitreous Body
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