CELLULAR COMMUNICATION IN MORPHOGENESIS

形态发生中的细胞通讯

• 批准号: 2185165
• 负责人: MARK A KRASNOW
• 金额: $ 12.28万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1991
• 资助国家: 美国
• 起止时间: 1991-08-09 至 1995-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2185165
• 关键词: Drosophilidae; anatomy; autosomal recessive trait; cell cell interaction; cell sorting; confocal scanning microscopy; developmental genetics; embryogenesis; fluoresceins; fluorescence microscopy; gene expression; gene mutation; genetic enhancer element; genetic regulation; histogenesis; invertebrate embryology; molecular cloning; monoclonal antibody; photoactivation; stainings; time resolved data; trachea

Very little is known of the means by which genes control the three- dimensional structure of the organs of an animal. In most screens for mutants affecting Drosophila melanogaster development, internal tissues have been ignored in favor of external tissues because they are not easily visualized. The objective of this proposal is to initiate a combined morphological, genetic, and biochemical study of the development of the Drosophila embryonic tracheal system, an elaborate but regular branched tubular structure which is the animal's respiratory organ. Formation of the tracheal system involves the invagination of a defined set of epithelial cells, the tracheal precursor cells, in each thoracic and abdominal segment to form blind-ended tubes; branching of the blind end and branch outgrowth; fusion of branches from adjacent segments to form a continuous tube; and further branching and tissue penetration by these secondary branches. The long-term goals of this research are to identify the gene products that control these morphogenetic processes, focusing on the biochemical mechanisms of branch formation and the outgrowth and fusion of the branches. The specific aims of this proposal are: 1. To characterize the normal growth and development of the embryonic tracheal system by fluorescence microscopy and high resolution confocal microscopy using a monoclonal antibody that stains trachea throughout embryogenesis. 2. To elucidate dynamic aspects of tracheal tube formation, branching, and branch outgrowth and fusion by real-time imaging of tracheal development after photo-activation of a caged fluorescein molecule in tracheal precursor cells. 3. To isolate genes involved in tracheal development, by screening for recessive mutations that alter tracheal anatomy and by screening lacZ enhancer-detector strains to identify genes expressed in or near the developing trachea. 4. To initiate the molecular cloning and molecular genetic characterization of one or more of the identified genes involved in tracheal development. 5. To purify tracheal precursor cells by a new approach called whole animal cell sorting to establish an in vitro system for tracheal development.

对基因控制三个的手段知之甚少 动物器官的尺寸结构。 在大多数屏幕中 影响果蝇的突变体，内部组织 由于不容易而被忽略以支持外部组织 可视化。 该提议的目的是启动一个合并 形态学，遗传和生化研究 果蝇胚胎气管系统，一种精致但规则的分支 管状结构是动物的呼吸器官。 形成 气管系统涉及一组定义的一组 上皮细胞，气管前体细胞，每个胸部和 腹部段形成盲管；盲目的分支和 分支出口；从相邻段的分支融合以形成A 连续管；以及进一步的分支和组织穿透 次级分支。 这项研究的长期目标是确定 控制这些形态发生过程的基因产品，重点是 分支形成的生化机制以及产物和融合 分支。 该提案的具体目的是： 1。表征胚胎的正常生长和发展 气管系统通过荧光显微镜和高分辨率共焦 使用单克隆抗体的显微镜，该抗体在整个过程中染色气管 胚胎发生。 2。阐明气管管形成，分支和分支的动态方面 通过气管开发的实时成像进行分支的生长和融合 光激活气管中的笼荧光素分子 前体细胞。 3。通过筛选涉及气管发育的基因 隐性突变，改变气管解剖结构并筛选LACZ 增强子检测器菌株以鉴定在或附近表达的基因 发展气管。 4。启动分子克隆和分子遗传表征 一个或多个涉及气管发育的鉴定基因。 5。通过一种称为全动物的新方法净化气管前体细胞 细胞分类以建立用于气管发育的体外系统。