MOLECULAR GENETICS OF LEARNING AND MEMORY IN DROSOPHILA

描述：研究人员正在进行分子表征 四个新的果蝇基因，latheo，linotte，nalyot和goolovan，都是 在P元件(转座子)插入突变中被鉴定 用于在巴甫洛夫嗅觉调节后影响3小时记忆的突变体。 对latheo、linotte和nalyot突变体的行为分析表明 学习被打乱(Golovan突变体还没有被测试)，而每个 突变体对气味和电击的感知和反应能力出现 很正常。 利用这些基因是P元素标记的分子 基因分析已经开始。Latheo和linotte转录单位 都被鉴定出来，并且每个都编码一种新的蛋白质。完全解救 已在成人转基因车床上实现学习/记忆缺陷 Linotte突变体，每个携带其各自的可诱导构建体 野生型序列。基因组DNA侧翼和Nalyot与Golovan P元件 插入片段已被回收并测序。这两种产品的比较 数据库的序列显示，nalyot P元件是 插入到ADF-1基因的内含子中，Golovan P元件是 插入了70个BP 5‘额外的巨毛虫(EMC)。申请者建议 通过追求以下几点来扩展我们对这些基因的描述： 1.突变型和突变型嗅觉学习和记忆的特征 转基因果蝇 2.转录本的分子特征及基因组调控 这些基因的区域 3.RNA和蛋白质表达模式的原位表征 4.使用双突变对基因相互作用进行表征， 免疫细胞学共定位、酵母双杂交选择和 差异化屏幕。 考虑到学习和学习的行为特性的广泛同源性 对于果蝇的记忆和参与学习的基因，其他 无脊椎动物和脊椎动物，申请者期望从 他们对果蝇的研究有助于我们理解 学习和记忆的分子机制。最终是这样的 信息将被用于开发新的诊断工具、药物 以及人类认知功能障碍的基因疗法。

DESCRIPTION: The investigators are pursueing the molecular characterization of four new Drosophila genes, latheo, linotte, nalyot and golovan, all of which were identified in a P element (transposon) insertional mutagenesis for mutants affecting 3-hr memory after Pavlovian olfactory conditioning. Behavioral analyses of latheo, linotte and nalyot mutants suggest that learning is disrupted (golovan mutants have not yet been tested), while each mutant's abilities to sense, and react to, odors and electric shock appear normal. Capitalizing on the fact that these genes were P element-tagged, molecular genetic analyses have begun. The latheo and linotte transcription units have been identified and each encodes a novel protein. Complete rescue of the leaning/memory defects has been achieved in adult transgenic latheo and linotte mutants, each carrying inducible constructs of their respective wild-type sequences. Genomic DNA flanking and nalyot and golovan P element insertions has been plasmid rescued and sequenced. Comparisons of these sequences to the databases have revealed that the nalyot P element is inserted in an intron of the Adf-1 gene and that the golovan P element is inserted 70 bp 5' of extra macrochaetae (emc). The applicants propose to extend our characterizations of these genes by pursuing the following: 1. Characterization of olfactory leaning and memory in mutant and transgenic flies 2. Molecular characterization of the transcripts and genomic regulatory regions of these genes 3. In situ characterizations of RNA and protein patterns of expressions 4. Characterization of gene interactions using double-mutants, immunocytological co-localization, yeast two-hybrid selection and differential screens. Given the extensive homologies for behavioral properties of learning and memory and for genes involved with learning among fruit flies, other invertebrates and vertebrates, the applicants expect knowledge gained from their studies of fruit flies to contribute to our understanding of the molecular mechanisms underlying learning and memory. Ultimately such information will be used to develop new diagnostic tools, pharmaceuticals and gene therapies for cognitive dysfunction in humans.