Construction and stability of photoreceptor outer segment discs

感光器外节盘的结构和稳定性

• 批准号: 10091444
• 负责人: Peter Deane Calvert
• 金额: $ 42.88万
• 依托单位: UPSTATE MEDICAL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-04-01 至 2023-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10091444
• 关键词: 11 cis Retinal; Address; Adhesions; Binding Sites; Biochemical; Biological; Biology; Biophysical Process; Biophysics; Blindness; Cells; Cellular biology; Charge; Cone; Defect; Dimerization; Disease; Divalent Cations; Electrolytes; Electrostatics; Elements; Environment; Eye; Face; Genes; Health; Imaging Device; Ions; Lead; Light; Maintenance; Membrane; Membrane Proteins; Molecular; Molecular Genetics; Monovalent Cations; Morphology; Mutation; Opsin; Organelles; Orphan; Photons; Photoreceptors; Phototransduction; Physiological; Physiology; Process; Proteins; Retinal Cone; Retinal Degeneration; Retinal Photoreceptors; Retinitis Pigmentosa; Rhodopsin; Rod; Role; Signal Transduction Pathway; Structure; Techniques; Transgenic Organisms; Vertebrate Photoreceptors; Vision; Work; Xenopus; cilium biogenesis; dimer; extracellular; fluorescence imaging; glycosylation; nanometer; novel therapeutics; photoreceptor degeneration; photoreceptor disc; protein folding; retinal rods; sight restoration; trafficking; virtual

Project summary The objectives of this proposal are to determine the mechanisms of photoreceptor disc membrane elaboration. Disc membranes are the central organelles for vision where photons are absorbed and phototransduction ensues. Devastating blinding disease manifest as disruptions in orderly stacks of outer segment discs, showing that maintaining this order is essential to photoreceptor health and function. A key element to disc formation is the close juxtaposition of the intradiscal membrane faces, which are only a few nanometers apart. This apposition is highly energetically unfavorable and thus produces an energy barrier that must be overcome for discs to form. Finding out how this energy barrier is overcome is important for understanding rod and cone outer segment elaboration and stability. We will use state of the art live cell fluorescence imaging tools developed in our lab, powerful transgenic and gene editing techniques in Xenopus and biochemical and cell biological approaches to address the following aims: Aim 1: Determine the roles of opsin dimers or higher order oligomers in forming and maintaining rod and cone photoreceptor discs. Aim 2: Determine the roles of opsin N-glycosylation in generating and maintaining rod and cone photoreceptor discs. Aim 3: Determine the contributions of membrane charge shielding by electrolytes to rod and cone disc formation and stability.

项目摘要 本提案的目的是确定感光盘膜的机制 详细说明。盘膜是视觉的中心细胞器，光子在这里被吸收 和光传导增强。毁灭性的致盲疾病表现为 堆叠的外节盘，表明维持这种秩序是必不可少的感光器 健康和功能。椎间盘形成的一个关键因素是椎间盘内 膜面，这是只有几个纳米分开。这种并置是非常积极的 这是不利的，因此产生了形成盘必须克服的能量势垒。找到 如何克服这个能量障碍是重要的了解棒和锥外 段细化和稳定性。我们将使用最先进的活细胞荧光成像工具 在我们的实验室开发的，强大的转基因和基因编辑技术在非洲爪蟾， 生物化学和细胞生物学方法，以实现以下目标： 目的1：确定视蛋白二聚体或更高级的寡聚体在形成和维持视蛋白功能中的作用。 视杆和视锥感光盘。 目的2：研究视蛋白N-糖基化在视杆细胞和视锥细胞形成和维持中的作用 感光盘 目的3：确定电解质对杆和锥的膜电荷屏蔽的贡献 椎间盘形成和稳定性。