CELL COMMUNICATION IN RETINAL MORPHOGENESIS

拟议研究的长期目标是理解这部小说 果蝇眼睛发育过程中细胞通讯途径的揭示 由脂肪分面(Faf)基因决定。FAF在细胞相互作用中起着关键作用 防止特定细胞出现在眼睛的早期阶段 从不适当地成为光感受器发展而来，因此 扰乱正常视网膜功能所需的对称性。一个 分子和遗传方法的结合将被用来回答 四个具体的问题。 1.Faf蛋白在哪里表达和亚细胞定位？ 将产生抗Faf抗体和标记表位的Faf蛋白，并 用于检测双眼发育期Faf蛋白的表达 和电子显微镜。 2.在眼睛发育过程中何时需要Faf功能，其中 细胞？ 异源启动子将被用来表达Faf。 在发育过程中不同时间发育的眼睛，以及对 评估视网膜形态发生。 3.Faf蛋白有哪些功能结构域？ Faf蛋白在进化上的保守结构域将通过 果蝇Faf基因的分离及DNA测定 ITS编码区序列。此外，许多突变体的DNA序列也发生了变化 FAF等位基因将被鉴定出来。由此确定的FAF蛋白结构域将 在转基因果蝇身上进行功能测试。 4.在细胞通讯通路中有哪些组成部分 FAF函数？ 将对果蝇进行基因筛查，以寻找新的突变 增强或抑制Faf突变果蝇的眼睛缺陷。一块屏幕 酵母中Faf蛋白与蛋白质间直接相互作用的研究 结构域和其他眼部蛋白将被执行。 在脊椎动物和果蝇中，使用同源蛋白质进行细胞- 细胞相互作用是司空见惯的。因此，虽然解剖学上的 果蝇和脊椎动物的眼睛有很大的不同，利用果蝇 作为一个模型系统将允许发现细胞通信 这两种机制很可能是共同的。

The long-term goal of the proposed research is to understand the novel pathway of cell communication in the developing Drosophila eye revealed by the fat facets (faf) gene. faf plays a key role in cell interactions that prevent particular cells present in the earliest stages of eye development from inappropriately becoming photoreceptors, and thus disturbing the symmetry that proper retinal function requires. A combination of molecular and genetic approaches will be used to answer four specific questions. 1. Where is FAF protein expressed and subcellularly localized? Anti-FAF antibodies and epitope-tagged FAF proteins will be generated and used to examine FAF protein expression in developing eyes with both light and electron microscopy. 2. When during eye development is faf function required and in which cells? Heterologous promoters will be used to express faf in portions of the developing eye at different times during development, and the effects on retinal morphogenesis assessed. 3. What are the functional domains of FAF protein? Evolutionarily conserved domains of FAF protein will be identified by isolating the Drosophila virilis faf gene and determining the DNA sequence of its coding region. Also, DNA sequence changes in many mutant faf alleles will be identified. FAF protein domains thus identified will be tested for function in transgenic flies. 4. What are the components of the cell communication pathway in which faf functions? A genetic screen in Drosophila will be conducted to find novel mutations that enhance or suppress the eye defects in faf mutant flies. A screen in yeast for direct protein-protein interactions between FAF protein domains and other eye proteins will be performed. In vertebrates and Drosophila, the use of homologous proteins for cell- cell interactions is commonplace. Thus, although the anatomy of the Drosophila and vertebrate eyes are quite different, the use of Drosophila as a model system will allow the discovery of cell communication mechanisms likely to be common to both.