Astrocyte Development and Reactive Gliosis

星形胶质细胞发育和反应性神经胶质增生

基本信息

批准号：
7941814
负责人：
TERI L BELECKY-ADAMS
金额：
$ 30.02万
依托单位：
INDIANA UNIV-PURDUE UNIV AT INDIANAPOLIS
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-09-30 至 2013-09-29
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7941814
关键词：
Age related macular degeneration Antisense Technology Astrocytes Binding Biological Assay Biology Blood - brain barrier anatomy Cell Count Cells Co-Immunoprecipitations DNA-Binding Proteins Data Development Diabetic Retinopathy Disease Electrophoretic Mobility Shift Assay Electroporation Environment Erinaceidae Eye Eye Injuries Eye diseases Funding GLI2 gene Gene Expression Regulation Glaucoma Gliosis Goals Growth Factor Human Image Analysis Immunohistochemistry In Situ Hybridization In Vitro Injury Laboratories Lead Luciferases Mus Natural regeneration Nervous system structure Neuroglia Neurons Nuclear Orphan Receptor Nutrient Optic Nerve Optic Nerve Injuries Pathway interactions Play Polymerase Chain Reaction Process Property Proteins Ranvier&apos s Nodes Regulation Repression Repressor Proteins Retina Retinal Retinal Ganglion Cells Retinitis Pigmentosa Retinopathy of Prematurity Reverse Transcriptase Polymerase Chain Reaction Role Signal Pathway Signal Transduction Sonic Hedgehog Pathway Testing Tetracyclines Up-Regulation Visual system structure Western Blotting bone morphogenetic protein 7 cell transformation chromatin immunoprecipitation diencephalon ganglion cell immunocytochemistry in vitro Assay in vivo inhibitor/antagonist injured ionic balance loss of function member novel optic cup optic nerve regeneration optic stalk overexpression progenitor promoter public health relevance research study response to injury therapeutic target transcription factor vector

项目摘要

DESCRIPTION (provided by applicant): The long term goal of this laboratory is to enhance the ability of the human nervous system, particularly the retina and/or optic nerve, to regenerate. This proposal, if funded, would determine the role of two factors, bone morphogenetic protein 7 (BMP7) and sonic hedgehog (SHH), in increasing the development of a particularly detrimental cell state known as reactive gliosis. Reactive glia are present in many eye diseases, including glaucoma, age-related macular degeneration, retinitis pigmentosa, diabetic retinopathy and retinopathy of prematurity. The cells that transform into reactive glia arise from the region that becomes the optic nerve, and in the normal uninjured or non-diseased eye, the astrocytes perform essential supportive functions for the ganglion cell neurons of the retina. However, upon injury to or in the diseased eye, these cells become reactive, whereupon they begin expressing molecules that are unfavorable to the regeneration of the optic nerve. This proposal focuses on determining the mechanism(s) whereby BMP7 and SHH regulate the expression of PAX2, a DNA- binding protein that is essential for the development of optic nerve astrocytes. This application proposes a novel mechanism in which both SHH and BMP7 modulate the expression of Pax2 during development by altering the association of an inhibitor of PAX2 expression. This application will test if this novel mechanism drives the development of reactive astrocytes in vitro and in vivo. The studies proposed in this application use a multi-disciplinary approach, including 1) in vitro assays to determine interactions of BMP and SHH pathway members with other regulatory DNA-binding proteins such as co-immunoprecipitation, chromatin immunoprecipitation, and luciferase assays, 2) in vitro and in vivo manipulations of BMP pathways following optic nerve injury to determine the role of BMP7 in reactive gliosis, 3) in vivo and in vitro antisense technology to test BMP7 as potential target for reducing reactive astrocytes, and 4) characterization of the phenotypic properties of optic nerve astrocytes in vitro and in vivo using immunohistochemistry, Western blots, in situ hybridization, quantitative polymerase chain reaction and image analysis. Information derived from these studies will further our understanding of the mechanisms necessary for the development of reactive astrocytes and may also provide therapeutic targets for decreasing reactive gliosis in optic nerve injury and disease. PUBLIC HEALTH RELEVANCE: The focus of this proposal is the study of the astrocytes in the optic nerve and their response to injury. By studying the role of growth factors in the regulation of genes that are necessary for the development of normal and reactive astrocytes, we hope to increase the therapeutic targets for diseases such as glaucoma, diabetic retinopathy, retinopathy of prematurity, age-related macular degeneration, and retinitis pigmentosa.

描述（由申请人提供）：该实验室的长期目标是增强人类神经系统，特别是视网膜和/或视神经的再生能力。该提案如果获得资助，将确定骨形态发生蛋白 7 (BMP7) 和音刺猬 (SHH) 这两个因素在促进一种特别有害的细胞状态（称为反应性神经胶质增生）的发展中的作用。反应性神经胶质细胞存在于许多眼部疾病中，包括青光眼、年龄相关性黄斑变性、色素性视网膜炎、糖尿病性视网膜病变和早产儿视网膜病变。转化为反应性神经胶质细胞的细胞来自成为视神经的区域，在正常的未受伤或未患病的眼睛中，星形胶质细胞对视网膜的神经节细胞神经元发挥重要的支持功能。然而，当患病眼睛受伤或在患病眼睛中时，这些细胞就会变得活跃，于是它们开始表达不利于视神经再生的分子。该提案的重点是确定 BMP7 和 SHH 调节 PAX2 表达的机制，PAX2 是一种对视神经星形胶质细胞发育至关重要的 DNA 结合蛋白。该申请提出了一种新机制，其中 SHH 和 BMP7 通过改变 PAX2 表达抑制剂的关联来调节发育过程中 Pax2 的表达。该应用将测试这种新机制是否在体外和体内驱动反应性星形胶质细胞的发育。本申请中提出的研究采用多学科方法，包括 1) 体外测定，以确定 BMP 和 SHH 通路成员与其他调节性 DNA 结合蛋白的相互作用，例如免疫共沉淀、染色质免疫沉淀和荧光素酶测定，2)视神经损伤后 BMP 通路的体外和体内操作以确定 BMP7 在反应性神经胶质增生中的作用，3) 体内和体外反义技术测试 BMP7作为减少反应性星形胶质细胞的潜在靶标，4) 使用免疫组织化学、蛋白质印迹、原位杂交、定量聚合酶链反应和图像分析来表征视神经星形胶质细胞的体外和体内表型特性。从这些研究中获得的信息将进一步我们对反应性星形胶质细胞发育所需机制的理解，也可能为减少视神经损伤和疾病中的反应性神经胶质增生提供治疗靶点。公共健康相关性：该提案的重点是研究视神经中的星形胶质细胞及其对损伤的反应。通过研究生长因子在正常和反应性星形胶质细胞发育所必需的基因调节中的作用，我们希望增加青光眼、糖尿病性视网膜病变、早产儿视网膜病变、年龄相关性黄斑变性、和色素性视网膜炎。