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A Mizota, X Mo, H Negishi, A Yokoyama, T Oshitari, M Ikejiri, E Sato, E Adachi-Usami; Transfer BDNF Plasmid by Electroporation Supports the Survival of Axotomized RGCs in Adult Rat . Invest. Ophthalmol. Vis. Sci. 2002;43(13):754.
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Purpose: To investigate whether intravitreal injection of Brain-Derived Neurotrophic Factor (BDNF) gene into retinal ganglion cells (RGCs) with in vivo electroporation could promote the survival of axotomized RGCs in adult rat. Methods: Under deep anesthesia, 4 µl of mouse BDNF plasmid (2.5µg/µl) was intravitreally microinjected to adult rats with a 31 gauge needle. Immediately after the injection, in vivo electroporation was applied to the eye ball. BDNF protein expression was observed immunohistochemically. One week after the injection, optic nerve (ON) was transected to induce retinal ganglion cells (RGCs) damage. One week after ON transection, the effect of retarding apoptosis was detected by comparing TdT-dUTP terminal nick-end labeling (TUNEL) staining positive ratio in the RGC layer. 2, 4, 6 weeks after ON transection, the survival RGCs was evaluated by counting the number of RGCs which were retrogradely labeled by 1,1'-dioctadecyl-3, 3, 3', 3'-tetramethylindocarbocyanine percholorate (DiI) injected into superior colliculus. Results: The overexpression of BDNF could be detected by immunohistochemical staining till 5 weeks after the injection. BDNF gene injected eyes with in vivo electroporation had a higher rescue ratio in DiI staining and lower TUNEL positive cells ratio than the controls. Conclusion: . By BDNF gene transfer into RGCs by in vivo electroporation overexpression of BDNF protein could be detected and this overexpression of BDNF protein could promote the survival of RGCs from apoptosis. And also, in vivo electroporation was proved to be an effective method for direct gene delivery into RGCs.
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