PMID- 12091443 OWN - NLM STAT- MEDLINE DCOM- 20020715 LR - 20061115 IS - 0146-0404 (Print) IS - 0146-0404 (Linking) VI - 43 IP - 7 DP - 2002 Jul TI - Rescue of axotomized retinal ganglion cells by BDNF gene electroporation in adult rats. PG - 2401-5 AB - 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. FAU - Mo, Xiaofen AU - Mo X AD - 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 FAU - Yokoyama, Akiko AU - Yokoyama A FAU - Oshitari, Toshiyuki AU - Oshitari T FAU - Negishi, Hisanari AU - Negishi H FAU - Dezawa, Mari AU - Dezawa M FAU - Mizota, Atsushi AU - Mizota A FAU - Adachi-Usami, Emiko AU - Adachi-Usami E LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't PL - United States TA - Invest Ophthalmol Vis Sci JT - Investigative ophthalmology & visual science JID - 7703701 RN - 0 (Brain-Derived Neurotrophic Factor) SB - IM MH - Animals MH - *Apoptosis MH - Axotomy MH - Blotting, Western MH - Brain-Derived Neurotrophic Factor/*genetics/metabolism MH - Cell Count MH - Cell Survival/*physiology MH - Electroporation/*methods MH - Fluorescent Antibody Technique, Indirect MH - Gene Expression MH - Gene Transfer Techniques MH - Genetic Vectors MH - In Situ Nick-End Labeling MH - Male MH - Optic Nerve/surgery MH - Rats MH - Rats, Wistar MH - Retinal Ganglion Cells/*cytology/physiology MH - Transfection EDAT- 2002/07/02 10:00 MHDA- 2002/07/16 10:01 CRDT- 2002/07/02 10:00 PHST- 2002/07/02 10:00 [pubmed] PHST- 2002/07/16 10:01 [medline] PHST- 2002/07/02 10:00 [entrez] PST - ppublish SO - Invest Ophthalmol Vis Sci. 2002 Jul;43(7):2401-5.