Hangai M, Kaneda Y, Tanihara H, Honda Y
Department of Ophthalmology and Visual Science, Graduate School of Medicine, Kyoto University, Japan.
Invest Ophthalmol Vis Sci. 1996 Dec;37(13):2678-85.
To determine whether a reporter gene can be introduced into rat and mouse retinas in vivo using the authors' novel liposome system.
Cytomegalovirus-promoted LacZ genes and nonhistone nuclear protein, high mobility group 1 (HMG1), were coencapsulated in liposomes by agitation and sonication. The liposomes were then coated with the envelope of inactivated hemagglutinating virus of Japan (HVJ; Sendai virus) by fusion (HVJ liposome). The HVJ liposome solution was injected into the vitreous cavity of adult Sprague-Dawley rats (n = 30) or the subretinal space of adult CD-1 mice (n = 42). LacZ expression was assessed by beta-galactosidase assay.
LacZ expression was demonstrated in the photoreceptors as long as 30 days after intravitreal and subretinal injections. beta-Gal activity was observed also in neurons and glial cells in the ganglion cell layer of the intravitreally injected rats and, although at lower intensity, in the retinal pigment epithelium of both animals. No inflammation or toxic effects secondary to the HVJ liposomes were detected on histologic examination.
In vivo transfer and expression of a reporter gene into adult mammalian retina can be achieved using HVJ liposomes. This method offers a promise as a nonviral system for in vivo gene transfer into adult mammalian neural retina.
使用作者新的脂质体系统，确定报告基因能否在大鼠和小鼠视网膜内进行体内导入。
通过搅拌和超声处理，将巨细胞病毒启动的LacZ基因与非组蛋白核蛋白高迁移率族蛋白1（HMG1）共包封于脂质体中。然后通过融合（HVJ脂质体）用灭活的日本血凝病毒（HVJ；仙台病毒）包膜包被脂质体。将HVJ脂质体溶液注入成年Sprague-Dawley大鼠（n = 30）的玻璃体腔或成年CD-1小鼠（n = 42）的视网膜下间隙。通过β-半乳糖苷酶测定评估LacZ表达。
玻璃体内和视网膜下注射后长达30天，在光感受器中均显示有LacZ表达。在玻璃体内注射大鼠的神经节细胞层的神经元和神经胶质细胞中也观察到β-半乳糖苷酶活性，并且在两种动物的视网膜色素上皮中也观察到β-半乳糖苷酶活性，尽管强度较低。组织学检查未检测到继发于HVJ脂质体的炎症或毒性作用。
使用HVJ脂质体可实现报告基因在成年哺乳动物视网膜中的体内转移和表达。该方法有望成为一种用于成年哺乳动物神经视网膜体内基因转移的非病毒系统。
