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.

项目总结/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， 故障影响信息处理和细胞活力。然而，在这方面存在着根本性的差距。 了解生物钟如何控制健康和患病视网膜组织的功能通路。继续 这一差距的存在是一个重要的问题，因为在填补这一差距之前， 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发育和维持， 视网膜的视觉处理这个项目的目的是确定生物钟是如何在 光感受器（即视杆细胞、视锥细胞和opn4/表达黑视素的视网膜神经节细胞或ipRGC）控制视网膜神经节细胞的光感受性（即视杆细胞、视锥细胞和opn4/表达黑视素的视网膜神经节细胞或ipRGC）。 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， 通过受限的时钟路径运行。我们的中心假设是根据我们的 拥有该领域的初步数据和最新出版物。拟议研究的理由是， 基因沉默的时钟机制，特别是在感光细胞类型，我们将能够联系这一点， 细胞类型的时钟与视网膜发育和功能相关的不同时钟通路。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。我们将追捕两名 具体目的：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）识别候选基因和受光感受器时钟控制的信号通路。 根据第一个目标，视网膜组织的形态学分析和各种电生理和行为 方法将用于已经建立的条件性时钟缺陷小鼠系。在第二个目标下， 我们将结合联合收割机，锥状细胞、杆状细胞或ipRGC的单细胞辅助分选（FACS），RNA测序， 荧光RNA原位杂交（FISH）和免疫细胞化学。另外我们 已经制定了一项计划，将RNAseq数据分析优先考虑到一些特定的感兴趣的生物过程。 拟议的研究意义重大，因为它有望纵向推进和扩大对 生物钟如何控制视网膜发育和功能，并将提供关键的缺失信息 有关生物钟的路径。最终，这些知识有可能增加我们的 理解光感受器维持的一般规则和导致光感受器的事件。 它们在退行性疾病如年龄相关性黄斑变性中的功能障碍。