MEMBRANE BIOSYNTHESIS IN NORMAL AND DYSTROPHIC RETINAS

正常和营养不良视网膜中的膜生物合成

• 批准号: 2902573
• 负责人: DAVID S PAPERMASTER
• 金额: $ 37.69万
• 依托单位: UNIVERSITY OF CONNECTICUT SCH OF MED/DNT
• 依托单位国家: 美国
• 财政年份: 1986
• 资助国家: 美国
• 起止时间: 1986-07-01 至 2002-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2902573
• 关键词: Xenopus; apoptosis; cone cell; confocal scanning microscopy; electron microscopy; gene expression; gene mutation; genetically modified animals; green fluorescent proteins; membrane biogenesis; membrane proteins; model design /development; molecular shape; organ culture; protein biosynthesis; protein transport; retina; retina degeneration; rhodopsin; rod cell; subtraction hybridization; visual photoreceptor

DESCRIPTION (adapted from the applicant's abstract): The principal investigator seeks to link mutations that can induce retinal degeneration in humans with disordered rhodopsin biosynthesis/transport and photoreceptor cell death. Many human retinal degenerations involve mutant genes expressed only in rods, yet cones die. How the injury of rods leads to cone death is unclear. The specific aims of this application are to: (1) develop transgenic Xenopus laevis as a model system to determine factors that perturb rhodopsin synthesis and transport and (2) develop models of retinal degeneration in transgenic Xenopus laevis to permit assessment of the impact of the death of rods on cone survival. To accomplish this, the investigator will determine: (1) where the transgene products are localized within photoreceptor cells; (2) the effect of the transgene products on the biosynthesis and transport of outer segment membrane proteins; and (3) their impact on photoreceptor cell viability. Preliminary results have been obtained using the opsin promoter to drive expression of green fluorescent protein (GFP) and GFP fusion proteins. The transgene products are expressed only in the major rods and are detectable 5 days after fertilization in the eye of the living tadpole. Since the frog retina is cone-rich, these transgenic frogs are suitable for examining the interaction of dying rods and the neighboring cones as models of human retinitis pigmentosa.

描述（改编自申请人的摘要）：首席研究员 试图将可诱发人类视网膜变性的突变与 视紫红质生物合成/运输紊乱和感光细胞死亡。许多 人类视网膜变性涉及仅在视杆细胞中表达的突变基因，但 视锥细胞死亡。视杆细胞的损伤如何导致视锥细胞死亡尚不清楚。具体的 本申请的目的是：（1）开发转基因非洲爪蟾作为 模型系统以确定扰乱视紫红质合成的因素 运输和（2）开发转基因非洲爪蟾的视网膜变性模型 莱维斯允许评估视杆细胞死亡对视锥细胞的影响 生存。为了实现这一点，研究者将确定：（1） 转基因产物定位于感光细胞内； (2)效果 转基因产物对外节生物合成和运输的影响 膜蛋白； (3)它们对感光细胞活力的影响。 使用视蛋白启动子驱动获得了初步结果 绿色荧光蛋白（GFP）和GFP融合蛋白的表达。这 转基因产物仅在主要杆中表达并且可检测到 5 受精后几天，在活蝌蚪的眼睛里。自从青蛙 视网膜富含视锥细胞，这些转基因青蛙适合检查 垂死的视杆细胞和邻近视锥细胞的相互作用作为人类模型 视网膜色素变性。