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，lateo，linotte，nalyot和golovan， 这是在P元件（转座子）插入突变中鉴定的 巴甫洛夫嗅觉条件反射后影响3小时记忆的突变体。 对latheo，linotte和nalyot突变体的行为分析表明， 学习中断（Golovan突变体尚未测试），而每个 突变体的能力，感觉，并作出反应，气味和电击出现 正常 利用这些基因被P元素标记的事实， 基因分析已经开始。 latheo和linotte转录单位 已经被鉴定并且每一种都编码一种新的蛋白质。 彻底拯救 学习/记忆缺陷已经在成年转基因小鼠中实现， linotte突变体，每一个都携带它们各自的 野生型序列。 基因组DNA侧翼与nalyot和golovan P元件 插入物已被质粒拯救并测序。 这些比较 数据库的序列显示，nalyot P元素是 插入Adf-1基因的内含子中，并且Golovan P元件 插入70 bp的额外的巨毛类（emc）5'。 申请人建议 通过以下方式扩展我们对这些基因的描述： 1. 突变型和非突变型小鼠嗅觉学习和记忆的特征 转基因果蝇 2. 转录物的分子表征和基因组调控 这些基因的区域 3. RNA和蛋白质表达模式的原位表征 4. 使用双突变体表征基因相互作用， 免疫细胞学共定位、酵母双杂交选择和 差分屏幕。 考虑到学习行为特性的广泛同源性， 记忆和与果蝇学习有关的基因， 无脊椎动物和脊椎动物，申请人希望获得的知识， 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 最终这样 信息将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.