Molecular Genetic Analysis of Sclera Development and Degeneration

巩膜发育和变性的分子遗传学分析

• 批准号: 8954458
• 负责人: WILLIAM R JEFFERY
• 金额: $ 38万
• 依托单位: UNIV OF MARYLAND, COLLEGE PARK
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-08-01 至 2020-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8954458
• 关键词: Affect; Cephalic; Chromosome Mapping; Collection; Defect; Development; Developmental Process; Embryo; Exhibits; Experimental Genetics; Eye; Eye Part; Eye diseases; Fishes; Gene Expression Profile; Gene Mutation; Genes; Genetic; Genomic Segment; Goals; Growth; Hereditary Disease; Image; Left; Maps; Mediating; Methods; Modeling; Molecular; Molecular Genetics; Mutation; Neural Crest; Optics; Phenotype; Process; Processed Genes; Quantitative Trait Loci; Research; Retina; Role; Sclera; Sequence Analysis; Shapes; Structure of retinal pigment epithelium; Surface; System; Techniques; Tissues; Transplantation; Vision; Visual; base; blind; design; eye primordia; flexibility; gene function; genetic analysis; genetic approach; genetic manipulation; genetic resource; genome sequencing; genomic tools; lens; mutant; novel; optic cup; public health relevance; success; teleost

 DESCRIPTION (provided by applicant): The sclera is the fibrous elastic coat surrounding the eyeball that provides the structural support and flexibility critical for focusing an image on the retina. Despite its importance in optic development and disease, the molecular mechanisms of sclera development and degeneration are poorly understood. This proposal will fill this gap by capitalizing on novel opportunities for studying sclera development and degeneration in the teleost Astyanax mexicanus, a single species consisting of eyed surface (surface fish) and blind cave (cavefish) forms. In this species, eye tissue manipulation methods, genomic tools, and - most importantly - the existence of a powerful genetic approach, are available. Eye primordia are initially formed in cavefish but subsequently arrest and degenerate. The first cavefish tissue to degenerate is the lens, which subsequently affects development of other optic tissues including the sclera. Transplantation of a surface fish embryonic lens into a cavefish optic cup restores the normal sclera phenotype, showing that the lens functions in sclera development. Other eye tissues, namely the retinal pigment epithelium (RPE) and cranial neural crest (CNC), may also control aspects of sclera development. The overall goal of this project is to determine the molecular mechanisms of sclera development and degeneration. The first specific aim is to investigate the roles of the lens, RPE, and CNC as organizers of sclera development. This aim will use a combination of physical and genetic manipulations to reveal the contributions of these eye tissues in organizing the developing sclera. The second specific aim will identify the genes expressed in the lens, RPE, CNC, and/or the sclera itself that mediate normal and abnormal sclera development. These genes will be filtered for specifically and reduced to a manageable number by mapping to a set of specific genomic regions (QTL) responsible for the abnormal sclera phenotype. The third specific aim will determine the function of the sclera related genes by manipulating their expression in surface fish and cavefish and determining the effects on sclera development. The fourth specific aim will identify the mutations in genes responsible for sclera defects. This study is designed to reveal the mechanisms, genes, and mutations responsible for visual decay through effects on development of the sclera.

 描述（由申请人提供）：巩膜是围绕眼球的纤维弹性涂层，其提供对于将图像聚焦在视网膜上至关重要的结构支撑和柔性。尽管其在视神经发育和疾病中的重要性，巩膜发育和变性的分子机制知之甚少。这项建议将填补这一空白，利用新的机会，研究巩膜发育和退化的硬骨鱼Astyanax墨西哥，一个单一的物种组成的眼睛表面（表面鱼）和盲洞（洞穴鱼）的形式。在这个物种中，眼组织操作方法，基因组工具，以及-最重要的-强大的遗传方法的存在，是可用的。眼原基最初在洞穴鱼中形成，但随后停滞并退化。第一个退化的洞穴鱼组织是透镜，它随后影响包括巩膜在内的其他视组织的发育。将表面鱼胚胎透镜移植到cavefish视杯中恢复了正常的巩膜表型，表明透镜在巩膜发育中起作用。其他眼组织，即视网膜色素上皮（RPE）和颅神经嵴（CNC），也可以控制巩膜发育的各个方面。本项目的总体目标是确定巩膜发育和退化的分子机制。第一个具体的目的是调查的作用，透镜，视网膜色素上皮细胞，和数控作为组织者的巩膜的发展。这一目标将使用物理和遗传操作的组合来揭示这些眼组织在组织发育中的巩膜中的贡献。第二个具体目标将鉴定在透镜、RPE、CNC和/或巩膜本身中表达的介导正常和异常巩膜发育的基因。这些基因将被特异性地过滤，并通过映射到一组负责异常巩膜表型的特定基因组区域（QTL）而减少到可管理的数量。第三个具体目标是通过操纵巩膜相关基因在表层鱼和洞穴鱼中的表达并确定其对巩膜发育的影响来确定巩膜相关基因的功能。第四个具体目标将确定巩膜缺陷的基因突变。本研究旨在通过对巩膜发育的影响来揭示视觉衰退的机制、基因和突变。