ASTEROID GENE FUNCTION IN DROSOPHILA EYE DEVELOPMENT

小行星基因在果蝇眼睛发育中的功能

• 批准号: 3465889
• 负责人: MICHAEL A KOTARSKI
• 金额: $ 9.37万
• 依托单位: WEST VIRGINIA UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1991
• 资助国家: 美国
• 起止时间: 1991-08-01 至 1996-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3465889
• 关键词: Drosophilidae; antibody; biological models; cell differentiation; cellular pathology; complementary DNA; compound eye; computer assisted sequence analysis; electron microscopy; fusion gene; gene expression; gene mutation; genetic library; genetic transcription; growth /development; histogenesis; immunocytochemistry; molecular cloning; molecular pathology; monoclonal antibody; nucleic acid probes; nucleic acid sequence; phase contrast microscopy; protein sequence; protein structure function; restriction mapping; tissue mosaicism

The long-term objective of this research is to describe the mechanisms of development and cell differentiation at the molecular level. The Drosophila eye is chosen as the experimental system. This proposal outlines experiments aimed at deciphering the role of the asteroid gene in eye development. Asteroid cDNAs will be isolated and the DNA sequence of the coding regions of the gene determined. This sequence will provide the protein sequence of the asteroid gene product and identify its cellular function. The cellular defect in the eye caused by asteroid mutations will be examined by light microscopy of adult eye tissue sections to identify missing or aberrant cell types. The timing of the developmental mishap caused by the lack of asteroid function will be investigated by light microscopy of whole-mounted imaginal discs that have been stained with antibodies to neural epitopes or to specific gene products expressed in the cells of the eye. The specific cell types that are missing or develop aberrantly will be identified by electron microscopy of sectioned immunohistochemically stained discs. A genetic mosaic analysis will be done to determine if asteroid is cell autonomous and to determine in which cells of the eye the asteroid gene product is needed for normal development. Finally, an asteroid-LacZ fusion construct will be made in an expression vector and used to produce monoclonal antibodies to the asteroid protein. These antibodies will then be used to localize the asteroid protein in the cells of the eye. These experiments will determine the role of the asteroid gene in eye development and suggest mechanisms by which this role is carried out. This research will be used as a starting point to investigate how asteroid interacts with other gene products in an informational pathway to determine cell fate and how it functions in the development of the entire organism.

这项研究的长期目标是描述 发育和细胞分化的分子机制 等级。 选择果蝇眼睛作为实验系统。 这 提案概述了旨在破译其作用的实验 小行星基因在眼睛发育中的作用。 小行星 cDNA 将被分离出来，并且编码的 DNA 序列 确定的基因区域。 该序列将提供蛋白质 小行星基因产物的序列并鉴定其细胞功能。 小行星突变引起的眼睛细胞缺陷将是 通过光学显微镜检查成人眼组织切片以识别 缺失或异常的细胞类型。 发育障碍发生的时间 由于小行星功能缺失造成的，将通过光进行调查 已染色的整个成虫盘的显微镜检查 针对神经表位或特定基因产物的抗体 眼睛的细胞。 缺失或缺失的特定细胞类型 异常发育将通过切片的电子显微镜来识别 免疫组织化学染色的光盘。 遗传镶嵌分析将是 确定小行星是否具有细胞自主性并确定 眼睛的哪些细胞需要小行星基因产物来维持正常 发展。 最后，小行星-LacZ 融合结构将在 表达载体并用于产生单克隆抗体 小行星蛋白。 然后这些抗体将用于定位 眼睛细胞中的小行星蛋白。 这些实验将确定小行星基因在 眼睛发育并提出发挥这一作用的机制。 这项研究将作为研究如何 小行星与信息通路中的其他基因产物相互作用 确定细胞命运及其在细胞发育中的功能 整个有机体。