The role of circadian clocks in photoreceptor cell development, maintenance and function

生物钟在感光细胞发育、维持和功能中的作用

• 批准号: 9765320
• 负责人: Christophe P. Ribelayga
• 金额: $ 19.25万
• 依托单位: UNIVERSITY OF TEXAS HLTH SCI CTR HOUSTON
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-01 至 2021-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9765320
• 关键词: ARNTL gene; Affect; Age; Age related macular degeneration; Aging; Amaze; Animal Model; Animals; Architecture; Behavioral; Binding Proteins; Biological Process; Candidate Disease Gene; Carrier Proteins; Cell Separation; Cell Survival; Cell physiology; Cells; Circadian Dysregulation; Cone; Contrast Sensitivity; Data; Data Analyses; Defect; Degenerative Disorder; Development; Disease; Electrophysiology (science); Event; Fluorescence; Gene Expression Regulation; Gene Proteins; Genes; Goals; Grant; Impairment; In Situ Hybridization; Knock-out; Knowledge; Link; Maintenance; Masks; Modeling; Morphology; Motor Activity; Mus; Neural Retina; Pathway interactions; Phenotype; Photoreceptors; Photosensitivity; Physiology; Process; Proteins; Publications; RNA; RNA analysis; Reporting; Research; Retina; Retinal; Retinal Degeneration; Retinal Diseases; Retinal Ganglion Cells; Role; Signal Pathway; Signal Transduction; Structural Protein; The Sun; Time; Tissues; Vertebrate Photoreceptors; Vision; base; cell type; circadian; circadian pacemaker; differential expression; immunocytochemistry; information processing; insight; interest; light intensity; melanopsin; mouse model; mutant; neuron development; novel; protein transport; retinal rods; transcriptome; transcriptome sequencing; visual information; visual processing

PROJECT SUMMARY/ABSTRACT In the retina, many aspects of physiology and function are controlled by circadian (24-h) clocks, and clock malfunction impinges on information processing and cell viability. However, there is a fundamental gap in understanding how clocks control functional pathways in both healthy and diseased retinal tissue. Continued existence of this gap represents an important problem because, until it is filled, understanding of the mechanisms that link circadian clock malfunction and retinal disorders will remain largely incomprehensible. Our long-term goal is to better understand how circadian clocks control development and maintenance of visual processing in the retina. The objective of this project is to determine how circadian clocks within photoreceptors (i.e. rods, cones and opn4/melanopsin-expressing retinal ganglion cells or ipRGCs) control the development, maintenance and/or function of these cells. Our central hypothesis is that circadian clocks are present in most retinal cell types and each cell type's clock controls specific aspects of retinal development and function through a restricted clock pathway. Our central hypothesis has been formulated on the basis of our own preliminary data and recent publications in the field. The rationale for the proposed research is that by genetically silencing the clock mechanism specifically in a photoreceptor cell type, we will be able to link this cell type's clock to distinct clock pathways associated with retinal development and function. We have developed new genetically modified mouse lines and generated strong preliminary data. We will pursue two Specific Aims: 1) Characterize retina-specific and photoreceptor cell type-specific clock-deficient mouse models; and 2) Identify candidate genes and signaling pathways under the control of the photoreceptor clocks. Under the first aim, morphological analysis of retinal tissue and a variety of electrophysiological and behavioral approaches will be used in the conditional clock-deficient mouse lines already created. Under the second aim, we will combine Fluorescence-Assisted single-Cell Sorting (FACS) of cones, rods or ipRGCs, RNA sequencing and analysis, Fluorescence RNA In Situ Hybridization (FISH), and immuno-cytochemistry. In addition, we have established a plan to prioritize RNAseq data analysis to a few specific biological processes of interest. The proposed research is significant because it is expected to vertically advance and expand understanding of how circadian clocks control retinal development and function and will provide critical missing information about the clock pathways involved. Ultimately, such knowledge has the potential to increase our understanding of the general rules governing the maintenance of photoreceptors and of the events leading to their malfunction in degenerative diseases such as age-related macular degeneration.

项目概要/摘要 在视网膜中，生理和功能的许多方面都由昼夜节律（24 小时）时钟控制，并且生物钟 故障会影响信息处理和细胞活力。但存在根本性差距 了解时钟如何控制健康和患病视网膜组织的功能通路。持续 这一差距的存在是一个重要问题，因为在填补这一差距之前，对 将生物钟故障和视网膜疾病联系起来的机制在很大程度上仍然难以理解。 我们的长期目标是更好地了解生物钟如何控制生物钟的发育和维持 视网膜中的视觉处理。该项目的目标是确定生物钟如何 光感受器（即视杆细胞、视锥细胞和表达 opn4/黑视蛋白的视网膜神经节细胞或 ipRGC）控制 这些细胞的发育、维持和/或功能。我们的中心假设是生物钟 存在于大多数视网膜细胞类型中，每种细胞类型的时钟控制视网膜发育的特定方面 通过受限的时钟通路发挥作用。我们的中心假设是在我们的基础上提出的 自己的初步数据和该领域的最新出版物。拟议研究的基本原理是 通过基因沉默特定于感光细胞类型的时钟机制，我们将能够将其联系起来 细胞类型的时钟与视网膜发育和功能相关的不同时钟途径。我们有 开发了新的转基因小鼠品系并产生了强有力的初步数据。我们将追求两个 具体目标：1) 表征视网膜特异性和感光细胞类型特异性时钟缺陷小鼠 模型； 2) 识别光感受器时钟控制下的候选基因和信号通路。 在第一个目标下，视网膜组织的形态学分析以及各种电生理和行为学分析 方法将用于已经创建的条件时钟缺陷小鼠系。在第二个目标下， 我们将结合视锥细胞、视杆细胞或 ipRGC 的荧光辅助单细胞分选 (FACS)、RNA 测序 和分析、荧光 RNA 原位杂交 (FISH) 和免疫细胞化学。此外，我们 已经制定了一项计划，将 RNAseq 数据分析优先考虑到一些感兴趣的特定生物过程。 拟议的研究意义重大，因为它有望垂直推进和扩大对 生物钟如何控制视网膜发育和功能并提供关键的缺失信息 关于所涉及的时钟路径。最终，这些知识有可能增加我们的知识 了解光感受器维护的一般规则以及导致光感受器发生的事件 它们在退行性疾病（如年龄相关性黄斑变性）中的功能障碍。