ASTEROID GENE FUNCTION IN DROSOPHILA EYE DEVELOPMENT

这项研究的长期目标是描述 发育和细胞分化的分子机制 水平 选择果蝇眼作为实验系统。 这 该提案概述了旨在破译的作用， 小行星基因在眼睛发育中的作用 小行星的cDNA将被分离出来， 基因决定的区域。 这个序列将提供蛋白质 小行星基因产物的序列，并确定其细胞功能。 由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，以确定 缺失或异常的细胞类型。 发育中的不幸发生的时间 小行星缺乏功能所造成的问题将由光来调查 已染色的完整安装的成虫盘的显微镜检查 针对神经表位或针对在神经细胞中表达的特异性基因产物的抗体。 眼睛的细胞。 缺少的特定细胞类型或 将通过切片的电子显微镜鉴定异常发展 化学染色的圆盘。 基因嵌合体分析将在 以确定小行星是否是细胞自主的， 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，一个小行星-LacZ融合结构将在 表达载体，并用于产生抗 小行星蛋白 这些抗体将被用来定位 眼睛细胞中的小行星蛋白。 这些实验将确定小行星基因在 眼睛的发展，并提出了这一作用的机制。 这项研究将被用作一个起点，调查如何 小行星在信息通路中与其他基因产物相互作用 以确定细胞命运及其在发育过程中的作用， 整个有机体。

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.