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：
8511669
负责人：
Krzysztof Palczewski
金额：
$ 29.83万
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/8511669
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 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 transcriptome sequencing 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.

描述（由申请人提供）：脊椎动物的视网膜有两个主要组成部分：神经视网膜，它使用称为视杆细胞和视锥细胞的特殊感光细胞来收集光线并将这些信息传输到大脑；以及视网膜色素上皮细胞（RPE），它的作用是作为血液的一部分：视网膜屏障并执行多种重要功能。 RPE 通过提供血液中的代谢物、参与视觉发色团的再生以及通过吞噬作用定期去除感光细胞最旧的部分来维持视网膜健康。感光细胞通过从其基部不断产生新的外节，同时以昼夜节律的方式在尖端释放成熟的外节，从而保持几乎恒定的长度。因此，有丝分裂后的 RPE 细胞在一生中吞噬大量物质，处理光感受器生存和更新所需的细胞废物。该功能是维持视力所必需的，因为参与 RPE 吞噬作用的基因突变可能导致进行性视网膜变性。在这里，我们提出了一套定量分析来剖析 RPE 光感受器吞噬作用背后的遗传因素。首先，我们将采用下一代大规模并行 RNA 测序 (RNA-Seq) 来绘制整个昼夜节律时间线的小鼠眼睛转录组图谱。此外，我们将使用定量蛋白质组学方法来获得导致吞噬脱落光感受器外节的信号转导途径的更完整图像。最后，我们将在培养物中培养原代 RPE 细胞，并使用新开发的技术来辨别 RPE 吞噬过程中的第二信使信号传导。这些方法将深入了解负责 RPE 吞噬作用的复杂信号网络，并有可能提高我们对吞噬过程的总体理解。