GENES EXPRESSED DURING EARLY EYE DEVELOPMENT

早期眼睛发育过程中表达的基因

• 批准号: 2331663
• 负责人: Olof H Sundin
• 金额: $ 23.5万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1995
• 资助国家: 美国
• 起止时间: 1995-02-01 至 1999-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2331663
• 关键词: corneal epithelium; developmental genetics; developmental neurobiology; embryogenesis; eye; gene expression; genetic library; human genetic material tag; in situ hybridization; lens; molecular cloning; neurogenetics; nonmammalian vertebrate embryology; nucleic acid sequence; optic nerve; polymerase chain reaction

Formation of the eye begins during the initial stages of vertebrate embryogenesis, a period when many inherited or environmentally determined malformations affecting the eye have their origin. Our long term goal, a basic understanding of this developmental process is likely to provide considerable insight into the diagnosis, prevention and treatment of such disorders. Crucial to our understanding of this process is knowing the identity of genes selectively expressed in the cellular primordia which define the early eye. Such genes are strong candidates for the genes which govern eye morphology and the determination of its cell types. Human counterparts of these genes are also prime candidates for genes defective in inherited eye disorders. Less severe malfunctions of such genes are also of interest as possible risk factors contributing to more common diseases such as cataract and glaucoma. Currently, very few genes are known that are selectively expressed in the early optic vesicle and lens primordium. The optic vesicle, an outgrowth of the embryonic forebrain, gives rise to the optic nerve, retina, pigment epithelium, ciliary body and iris. The lens primordium region of the undifferentiated head ectoderm generates the lens as well as the corneal epithelium. Currently, the only known gene selectively expressed in these primordia is Pax-6, a transcriptional regulator essential to early development of the eye. Defects in one copy of the human Pax-6 gene cause aniridia, an inherited ocular malformation syndrome. The scope of this proposal is to isolate novel genes selectively expressed in the optic vesicle and lens primordium of the chick embryo. This will be accomplished by the micro-dissection of optic vesicles and lens primordia followed by the use of new techniques to generate cDNA libraries which represent the mRNA populations extracted from these small samples of tissue. Differential screening will identify genes expressed at higher levels in these early eye primordia, and further screening by in situ hybridization of whole embryos will be used to determine their detailed pattern of expression. For genes exhibiting distinctive expression patterns involving elements of the early eye, full length cDNA clones will be isolated and their sequence determined. This sequence information will provide clues to the cellular localization and function of the encoded protein, as will the preparation of antibodies specific for these proteins. Finally, homologous human genes will be isolated and then mapped to individual human chromosomes in order to provide a starting point for the identification of clinical disorders which might be associated with these genes.

眼睛的形成在脊椎动物的初始阶段开始 胚胎发生，许多遗传或环境确定的时期 影响眼睛的畸形具有起源。 我们的长期目标，一个 对这种发展过程的基本理解可能会提供 对这种诊断，预防和治疗的深入了解 疾病。 对于我们对这一过程的理解至关重要的是知道 在细胞原始中选择性表达的基因的身份 定义早期的眼睛。 这些基因是基因的强大候选者 控制眼睛形态及其细胞类型的确定。 这些基因的人类对应也是基因的主要候选者 遗传性眼部疾病有缺陷。 这样的严重故障 基因也引起了可能的风险因素的关注 常见疾病，例如白内障和青光眼。 目前，很少有基因在 早期视囊和镜头原始。 视线囊泡，产物 胚胎前脑的，产生视神经，视网膜，色素 上皮，睫状体和虹膜。 镜头原始区域 未分化的头外胚层产生镜头和角膜 上皮。 目前，唯一在这些基因中有选择地表达的基因 Primordia是PAX-6，是早期必不可少的转录调节剂 眼睛的发育。 人类pax-6基因的一个副本中的缺陷原因 Aniridia，一种遗传的眼畸形综合征。 该建议的范围是选择性地表达新型基因 在雏鸡胚胎的视力囊泡和晶状体中。 这会 可以通过微隔囊和镜头的微观划分来实现 Primordia，然后使用新技术生成cDNA库 代表从这些小样本中提取的mRNA种群 组织。 差异筛选将确定在较高的基因 这些早期眼睛的水平，并在原位进行进一步筛选 整个胚胎的杂交将用于确定其详细信息 表达模式。 对于表现出独特表达的基因 涉及早期元素的图案，全长cDNA克隆将 隔离并确定其序列。 此序列信息将 为编码的细胞定位和功能提供线索 蛋白质，对这些抗体的制备也会 蛋白质。 最后，同源人类基因将被隔离，然后 映射到单个人类染色体以提供起始 鉴定临床疾病的点 与这些基因相关。