The Role of TULP1 in Photoreceptor Cells

TULP1 在感光细胞中的作用

• 批准号: 7582298
• 负责人: STEPHANIE A HAGSTROM
• 金额: $ 30万
• 依托单位: CLEVELAND CLINIC LERNER COM-CWRU
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-04-01 至 2011-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7582298
• 关键词: Address; Affect; Affinity; Binding; Biochemical Pathway; Biological Models; Biology; C-terminal; Carrier Proteins; Cell membrane; Data; Defect; Development; Disease; Dynamin; Dynamin I; Engineering; Family; Family member; Foundations; Future; Genes; Genetic Predisposition to Disease; Human; Immunoelectron Microscopy; Inherited; Intracellular Transport; Knock-out; Knowledge; Light; Link; Messenger RNA; Modality; Molecular; Mus; Mutagenesis; Mutate; Mutation; Neurons; Pathologic; Pathway interactions; Phenotype; Photoreceptors; Physiological; Positioning Attribute; Process; Protein translocation; Proteins; Publishing; Retina; Retinal; Retinal Degeneration; Retinal Diseases; Retinitis Pigmentosa; Rhodopsin; Role; Signaling Protein; Synapses; Testing; Therapeutic; Vesicle; Wild Type Mouse; Work; base; design; early onset; extracellular; immunocytochemistry; mutant; photoreceptor degeneration; protein expression; protein protein interaction; protein transport; research study; trafficking; trans-Golgi Network

TULP1 is a gene we identified to cause autosomal recessive retinitis pigmentosa, a hereditary retinal degeneration blinding nearly 1 million people worldwide. The genetic etiology of RP is known for about 50% of cases; however, the biochemical pathways involved in causing the disease much less. The overall objectives of the proposed studies are to explore the physiologic function of TULP1 in the retina and to define the pathologic mechanism leading to photoreceptor degeneration associated with TULP1 mutations. The TULP family consists of four proteins of unknown function, two of which are linked to photoreceptor degeneration. The two specific aims of this application are designed to test the central hypothesis that TULP1 is a component of the molecular machinery involved in the directional translocation of proteins in photoreceptor cells. The first specific aim is to determine the role of TULP1 in photoreceptor transport pathways. This will be accomplished by using immunocytochemistry to determine whether outer segment proteins, intracellular transport proteins and synaptic proteins are mistargeted in tulpt-/- retinas. Immunoelectron microscopy will be performed to determine if the proteins that are incorrectly transported are cargo on the extracellular vesicles in tulpl-/- retinas. The second specific aim is to determine the function of the TULP1/Dynamin-1 interaction identified in photoreceptor cells. This will be done by identifying the functional domains that interact between TULP1 and Dynamin-1 and determining whether TULP1 mutations that cause RP alter the binding between the two proteins. Experiments are also proposed to generate and phenotype mice lacking Dynamin-1 in photoreceptor cells using Cre-loxP mutagenesis. Transport pathways will be evaluated in these mice as described in aim 1. Since little is known about TULP proteins, discovering information regarding the function of TULP proteins should provide knowledge about the pathways involved in photoreceptor degeneration. It is possible that this work could form the foundation for future studies aimed at evaluating therapeutic modalities that might slow, stop, or reverse the course of retinal degeneration.

TULP1 是我们确定的导致常染色体隐性视网膜色素变性的基因，这是一种遗传性视网膜病 退化导致全世界近一百万人失明。 RP 的遗传病因已知约 50% 案件数量；然而，引起该疾病的生化途径要少得多。整体 拟议研究的目的是探索 TULP1 在视网膜中的生理功能并 定义导致与 TULP1 突变相关的光感受器变性的病理机制。 TULP家族由四种功能未知的蛋白质组成，其中两种与光感受器有关 退化。 该应用程序的两个具体目标旨在测试 TULP1 是一个中心假设 参与光感受器中蛋白质定向易位的分子机器的组成部分 细胞。第一个具体目标是确定 TULP1 在光感受器运输途径中的作用。这将 通过使用免疫细胞化学来确定外段蛋白、细胞内蛋白是否 转运蛋白和突触蛋白在 tulpt-/- 视网膜中定位错误。免疫电子显微镜将 进行以确定错误运输的蛋白质是否是细胞外的货物 tulpl-/- 视网膜中的囊泡。第二个具体目标是确定 TULP1/Dynamin-1 的功能 感光细胞中发现的相互作用。这将通过识别功能域来完成 TULP1 和 Dynamin-1 之间的相互作用，并确定导致 RP 的 TULP1 突变是否会改变 两种蛋白质之间的结合。还建议进行实验来产生缺乏的小鼠并对其进行表型分析 使用 Cre-loxP 诱变的感光细胞中的 Dynamin-1。运输路径将在 这些小鼠如目标 1 中所述。 由于对 TULP 蛋白知之甚少，因此发现有关 TULP 蛋白功能的信息 应提供有关光感受器变性所涉及途径的知识。有可能的是 这项工作可以为未来的研究奠定基础，旨在评估可能的治疗方式 减缓、停止或逆转视网膜变性的进程。