Photoreceptor Renewal by Retinal Pigmented Epithelium Phagocytosis

视网膜色素上皮吞噬作用的光感受器更新

基本信息

批准号：
8330430
负责人：
Krzysztof Palczewski
金额：
$ 31.4万
依托单位：
CASE WESTERN RESERVE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-08-01 至 2015-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8330430
关键词：
Affect Biosensor Blindness Blood Blood Circulation Brain Cell Death Cell membrane Cells Circadian Rhythms Complex Cyclic AMP Distant Electrophoresis Event Eye Fluorescence Resonance Energy Transfer Focal Adhesions Fourier Transform Gel Gene Expression Gene Expression Profile Gene Mutation Gene Proteins Genes Genetic Health Histocompatibility Testing Housekeeping Immunoblotting Immunohistochemistry Ingestion Inositol Ions Laboratories Lead Learning Length Life Light Maintenance Maps Mass Spectrum Analysis Microarray Analysis Mitotic Monitor Mus Mutation Natural regeneration Neural Retina Nutrient Participant Pathway interactions Phagocytosis Phosphopeptides Phosphorylation Phosphotransferases Photoreceptors Play Process Production Protein Analysis Protein Tyrosine Kinase Proteins Proteomics RNA RNA Sequences Retina Retinal Retinal Degeneration Role Sampling Second Messenger Systems Signal Transduction Signal Transduction Pathway Solutions Staining method Stains Structure of retinal pigment epithelium Techniques Time TimeLine Tissues Vertebrate Photoreceptors Vision Visual base cellular imaging chromophore human MERTK protein immunocytochemistry improved insight macrophage mass spectrometer member neutrophil next generation protein expression protein protein interaction response sarcoma second messenger small molecule src-Family Kinases titanium dioxide transmission process tripolyphosphate two-dimensional wasting

项目摘要

DESCRIPTION (provided by applicant): The vertebrate retina has two main components, the neural retina, that employs specialized photoreceptor cells called rods and cones to collect light and transmit this information to the brain, and the retinal pigmented epithelium (RPE), which serves as part of the blood: retina barrier and performs several important functions. The RPE maintains retinal health by providing metabolites from the bloodstream, participating in the regeneration of the visual chromophore, and periodically removing the oldest portions of photoreceptor cells by phagocytosis. Photoreceptor cells maintain a virtually constant length by continuously generating new outer segments from their base while simultaneously releasing their mature outer segments at the tip in a circadian manner. Thus, post-mitotic RPE cells phagocytose an immense amount of material over a lifetime, disposing of cell waste required for photoreceptor survival and renewal. This function is required to maintain vision, as mutations in genes involved in RPE phagocytosis can lead to progressive retinal degeneration. Here, we propose a set of quantitative analyses to dissect the genetic factors underlying photoreceptor phagocytosis by the RPE. First, we will employ next generation massively parallel RNA-sequencing (RNA-Seq) to map the mouse eye transcriptome throughout the circadian timeline. Additionally, we will use quantitative proteomic approaches to gain a more complete picture of the signal transduction pathway leading to engulfment of shed photoreceptor outer segments. Lastly, we will grow primary RPE cells in culture and use newly developed techniques to discern second messenger signaling during RPE phagocytosis. These approaches will provide insights into the complex signaling networks responsible for RPE phagocytosis and potentially improve our understanding of phagocytic processes in general. PUBLIC HEALTH RELEVANCE: Retinal health and maintenance requires an intimate interaction between the neural retina and the retinal pigmented epithelium (RPE). This interaction provides nutrients and substrates to the neural retina, as well as a continuous process whereby the RPE engulfs the oldest portions of photoreceptors to permit their renewal. This study will increase our understanding of how RPE phagocytosis occurs and potentially provide answers to mechanistic questions pertaining to other phagocytic processes.

描述（由申请人提供）：脊椎动物视网膜具有两个主要成分，即神经视网膜，它们采用了专门的光感受器细胞，称为杆和锥体来收集光线并将其传递给大脑，而视网膜色素色上皮（RPE）是血液的一部分：视网膜障碍物：视网膜障碍物和执行多个重要的功能。 RPE通过从血液中提供代谢产物，参与视觉发色团的再生，并通过吞噬作用定期去除最古老的光感受器细胞部分。光感受器细胞通过连续从其底部产生新的外部段，同时以昼夜节律的方式释放其成熟的外部段来保持几乎恒定的长度。因此，有丝分裂后的RPE细胞在一生中吞噬了大量材料，即处理感光器生存和更新所需的细胞废物。需要此功能以保持视力，因为参与RPE吞噬作用的基因的突变会导致进行性视网膜变性。在这里，我们提出了一组定量分析，以通过RPE剖析感光受体吞噬作用的遗传因素。首先，我们将采用大规模平行的RNA测序（RNA-Seq）来绘制整个昼夜节律时间表的小鼠眼镜转录组。此外，我们将使用定量蛋白质组学方法来获得更完整的信号转导途径的图像，从而导致散落的感光体外部段吞没。最后，我们将在培养物中种植原发性RPE细胞，并使用新开发的技术来辨别RPE吞噬作用期间的第二信使信号传导。这些方法将提供有关负责RPE吞噬作用的复杂信号网络的见解，并有可能提高我们对吞噬过程的理解。公共卫生相关性：视网膜健康和维护需要神经视网膜与视网膜色素上皮（RPE）之间的紧密相互作用。这种相互作用为神经视网膜提供营养和底物，以及连续的过程，RPE吞噬了最古老的光感受器以允许其续订。这项研究将提高我们对RPE吞噬作用如何发生的理解，并有可能为与其他吞噬过程有关的机理问题提供答案。