活体电转化技术在新生小鼠视网膜中的应用

doi:10.3724/SP.J.1005.2014.1173

遗传 ›› 2014, Vol. 36 ›› Issue (11): 1173-1178.doi: 10.3724/SP.J.1005.2014.1173

活体电转化技术在新生小鼠视网膜中的应用

肖丽容, 陈大年, 闫乃红

四川大学华西医院眼科/眼科学研究室,成都 610041

收稿日期:2014-10-14 出版日期:2014-11-20 发布日期:2014-10-28
通讯作者: 闫乃红,博士,副教授,研究方向：医学遗传学。E-mail: yannaihong@126.com E-mail:lirongxiao@126.com
作者简介:肖丽容,学士,专业方向：医学技术学。
基金资助:
国家自然科学基金项目(编号：81371024)和四川省科技厅科技支撑项目(编号：2014SZ0030)资助

In vivo electroporation of newborn mouse retina

Lirong Xiao, Danian Chen, Naihong Yan

Ophthalmic Laboratories &
Department of Ophthalmology, West China Hospital, Sichuan University, Chengdu 610041, China

Received:2014-10-14 Online:2014-11-20 Published:2014-10-28

摘要/Abstract

摘要： 活体电转化技术是在高电压的脉冲作用下,瞬态增加细胞膜的渗透性从而将外源基因高效导入细胞的方法。与病毒载体等其他方法相比,活体电转化技术具有安全、高效、快速、稳定及应用范围广等优点,近年来在很多组织和器官中得到广泛使用,包括在眼科研究领域。文章介绍了活体电转化技术在新生小鼠视网膜中的应用,通过新生小鼠视网膜下注射的方法,经几次高电压的脉冲,将高浓度的绿色荧光蛋白表达质粒导入新生小鼠视网膜细胞内。通过冰冻切片观察绿色荧光蛋白在视网膜中的表达。结果表明绿色荧光蛋白在视网膜外核层高表达,证实了活体电转化技术可以将外源基因高效、快捷的导入视网膜,从而为研究视网膜发育及功能提供一种有效的手段。

关键词: 活体电转化, 新生小鼠, 视网膜, 冰冻切片

Abstract: In vivo electroporation is the process referred to a transient increase in the permeability of cell membranes upon high electric field pulses and delivery of engineered constructs into cells. Compared with the viral-mediated gene transfer system, the in vivo electroporation technique has several advantages, such as safe, high efficiency, rapid, stable and wide application. Thus, this technique has been widely used in the studies of many tissues or organs, including the eye. Here, we report the application of in vivo electroporation in the newborn mouse retina. DNA plasmid of GFP expression vector with high concentration was directly injected into the subretinal space of neonatal mouse pups. The DNA was then transfected into the retinal cells after several pulses of high voltage. Transfected GFP allowed clear visualization of cell morphologies in cryo-sections and the GFP was highly expressed in retinal outer nuclear layer. The results showed that this technique can effectively transfer the genes into retinal cells. In vivo electroporation will be a useful tool for the study of retinal development and function.

Key words: electroporation, newborn mouse, retina, cryo-sections

肖丽容, 陈大年, 闫乃红. 活体电转化技术在新生小鼠视网膜中的应用[J]. 遗传, 2014, 36(11): 1173-1178.

Lirong Xiao, Danian Chen, Naihong Yan. In vivo electroporation of newborn mouse retina[J]. HEREDITAS(Beijing), 2014, 36(11): 1173-1178.

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活体电转化技术在新生小鼠视网膜中的应用

In vivo electroporation of newborn mouse retina

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