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Effect of in vivo Electroporation on Rat Retina

体内电穿孔对大鼠视网膜的影响

基本信息

批准号：
15591843
负责人：
MIZOTA Atsushi
金额：
$ 1.86万
依托单位：
Juntendo University (2004)Chiba University (2003)
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2003
资助国家：
日本
起止时间：
2003 至 2004
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15591843/
关键词：
electroporation retina gene transfer ERG neuroprotection BDNF HSP アポトーシス

项目摘要

Purpose : We have used electroporation to transfer the heat shock protein and brain derived neurotorophic factor genes into retinal ganglion cells of rats. The purpose of this study was to determine the effect of electroporation on the rat retina as assessed by ERGs and morphological studies. Methods : Under deep anesthesia, right eye of 8-week-old Wistar rats was grasped between forcep-type electrodes, with the cathode attached to the corneal and anode attached to scleral surface Single electric pulse with electric field strengths of 10, 20, 30, 40, and 50V/cm were delivered. The pulse was square wave modulated, and the duration was 99 ms. ERGs were recorded before and one week after the electroporation. The rats were sacrificed after the ERGs, and the eyes were enucleated for histopathological study. The number of retinal ganglion cells were counted, and anterior segments were observed by microscopy. Results : Corneas of the rat that underwent electroporation with electric field strength of >30 V/cm showed thermal burns. Electric field strength of >20 V/cm, the amplitude of both a- and b-wave on ERGs and the number of retinal ganglion cells decreased significantly. Conclusions : These findings suggest that electroporation with an electric field strength of 10 V/cm or less does not affect the retina and cornea significantly in rats.

目的：我们采用电穿孔法将热休克蛋白和脑源性神经营养因子基因导入大鼠视网膜神经节细胞。本研究的目的是通过ERG和形态学研究来确定电穿孔对大鼠视网膜的影响。研究方法：在深度麻醉下，将8周龄Wistar大鼠的右眼夹持在钳型电极之间，其中阴极附接到角膜，阳极附接到巩膜表面。施加电场强度为10、20、30、40和50 V/cm的单次电脉冲。脉冲为方波调制，持续时间为99 ms。记录电穿孔前和电穿孔后一周的ERG。ERG检查后处死大鼠，摘除眼睛进行组织病理学研究。计数视网膜神经节细胞数，光镜下观察眼前节。结果：电场强度>30 V/cm的电穿孔大鼠角膜出现热灼伤。电场强度>20 V/cm时，视网膜电图a波和b波振幅及视网膜神经节细胞数均明显减少。结论：这些发现表明，电场强度为10 V/cm或更低的电穿孔不会显著影响大鼠的视网膜和角膜。