GENES FOR AXON GUIDANCE IN THE DROSOPHILA VISUAL SYSTEM

描述（改编自申请人的摘要）：广泛的，长期的目标， 本申请旨在研究细胞和分子机制， 参与建立视网膜之间精确的轴突连接 果蝇脑内光感受器神经元及其靶神经元。的 具体目标是：（1）使用遗传镶嵌分析来确定哪个细胞 视觉系统中的群体需要特定基因的功能;（2） 使用细胞类型特异性标记来确定哪些细胞群是 在突变动物中正常存在和分化;（3）表征 在分子水平上，根据在 目标1和2。嵌合分析将用于确定细胞群， 需要一个特定基因的功能， 将使用酵母FLP重组酶/FLP系统。识别特异性抗体 细胞类型将用于确定突变动物中的细胞身份。在 此外，具有用报告基因标记的特定细胞群的突变株 基因lacZ将被创建，不同的细胞类型将由一个 抗β-半乳糖苷酶抗体。分析将包括共聚焦 显微镜在这些分析中鉴定的基因将被表征为： 分子克隆，其中果蝇cDNA文库将使用 来自质粒拯救的探针和获得的突变基因的全长序列。 最后，将通过mRNA注射进行表型挽救分析 来证实克隆基因的拯救能力。

DESCRIPTION (Adapted from applicant's abstract): The broad, long-term goal of this application is to investigate the cellular and molecular mechanisms involved in establishing the precise axon connections between retinal photoreceptor neurons and their target neurons in the brain of Drosophila. The specific aims are: (1) to use genetic mosaic analysis to determine which cell populations in the visual system require the function of a particular gene; (2) to use cell-type specific markers to determine which cell populations are present and differentiate normally in the mutant animals; (3) to characterize at the molecular level genes selected on the basis of the analyses described in aims 1 and 2. Mosaic analysis will be used to determine cell populations that require the function of a particular gene, and the generation of mosaic animals will use the yeast FLP recombinase/FLP system. Antibodies recognizing specific cell types will be used to determine cell identity in mutant animals. In addition, mutant strains with specific cell populations marked with a reporter gene lacZ will be created and different cell types determined by an anti-beta-galactosidase antibody. The analysis will include confocal microscopy. The genes identified in these analyses will be characterized by molecular cloning, where Drosophila cDNA libraries will be screened using probes from plasmid rescue, and full-length sequences of mutant genes obtained. Finally, phenotype-rescuing analysis through mRNA injection will be performed to confirm the rescuing ability of a cloned gene.