CELL COMMUNICATION IN RETINAL MORPHOGENESIS

这项研究的长期目标是理解这部小说 果蝇眼睛发育中的细胞通讯途径揭示了 脂肪小面（FAF）基因。 faf在细胞相互作用中起关键作用 阻止眼睛早期出现的特定细胞 从不适当地成为光感受器的发展， 破坏了正常视网膜功能所需的对称性。 一 分子和遗传学方法的结合将被用来回答 四个具体问题。 1. FAF蛋白在哪里表达和亚细胞定位？ 将产生抗FAF抗体和表位标记的FAF蛋白， 用于检测在两种光下发育中的眼睛中FAF蛋白表达 和电子显微镜。 2. 在眼睛发育期间，何时需要faf功能， 细胞？ 异源启动子将被用于表达FAF的一部分， 在发育过程中的不同时期， 评估视网膜形态发生。 3. FAF蛋白的功能结构域有哪些？ FAF蛋白的进化保守结构域将通过以下方法鉴定： 分离果蝇faf基因， 其编码区的序列。 此外，许多突变体中的DNA序列变化 将鉴定FAF等位基因。 如此鉴定的FAF蛋白结构域将 在转基因果蝇中进行功能测试。 4. 细胞通讯途径的组成部分是什么， faf函数？ 将在果蝇中进行遗传筛选以发现新的突变 增强或抑制FAF突变果蝇的眼睛缺陷。 屏幕 在酵母中用于FAF蛋白之间的直接蛋白质-蛋白质相互作用 结构域和其他眼蛋白的研究。 在脊椎动物和果蝇中，使用同源蛋白质进行细胞- 细胞间的相互作用是常见的。 因此，虽然解剖学的 果蝇和脊椎动物的眼睛有很大的不同，果蝇使用 作为一个模型系统， 可能是两者共同的机制。

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.