Gene transfer into retinal ganglion cells by in vivo electroporation: a new approach.
We developed a new in vivo electroporation method to deliver genes into retinal ganglion cells (RGCs). Efficiency and degree of tissue damage were evaluated using green fluorescent protein (GFP) gene and TUNEL. Soon after the intravitreous injection of the GFP gene, electroporation (five electric pulses of 99 ms duration each and 12V/cm delivered twice 5 min apart) was carried out on the adult rat eyeball with the aid of tweezer-type disc electrodes attached to corneal (cathode) and scleral (anode) surfaces. GFP expression, exhibiting a maximum on day 7, was detectable for up to 21 days. DiI retrograde labeling of RGCs showed that 41.5% of the total ganglion cells in the electroinjected area were GFP-positive. Therefore, this new method may be a useful tool for the delivery of genes into RGCs.
Dezawa
M
M
Department of Anatomy, Yokohama City University School of Medicine, 3-9 Fukuura, Kanazawa-ku, 236-0004, Yokohama, Japan. dezawa@med.yokohama-cu.ac.jp
Takano
M
M
Negishi
H
H
Mo
X
X
Oshitari
T
T
Sawada
H
H
D000818:Animals; D018274:Electroporation / Q000379:methods*; D018014:Gene Transfer Techniques; D049452:Green Fluorescent Proteins; D006801:Humans; D020287:In Situ Nick-End Labeling; D008164:Luminescent Proteins / Q000235:genetics* / Q000378:metabolism; D018613:Microscopy, Confocal; D051381:Rats; D012160:Retina / Q000648:ultrastructure; D012165:Retinal Ganglion Cells
D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins