Functional Genomics of Drosophila Olfactory Memory

果蝇嗅觉记忆的功能基因组学

• 批准号: 6679677
• 负责人: TIM P TULLY
• 金额: $ 37.91万
• 依托单位: COLD SPRING HARBOR LABORATORY
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-08-01 至 2007-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6679677
• 关键词: Drosophilidae; arthropod genetics; developmental genetics; functional /structural genomics; genetic transcription; laboratory rabbit; long term memory; microarray technology; neurogenetics; olfactory stimulus

DESCRIPTION (provided by applicant): The molecular basis of LTM formation appears to be evolutionarily conserved. In particular, simple forms of learning appear to reflect the conserved cellular functions of neurons, while more complex behavioral tasks reflect the diversity of neuronal systems (anatomy) across species. Nonetheless, memory of these complex tasks appears to invoke a common underlying synaptic plasticity. Consequently, we have capitalized on the economy-of-scale and the availability of genetic tools in Drosophila to focus on the discovery of the downstream genes involved in LTM Via a combination of DNA microarrays and a behavioral screen for single-gene mutants with defective memory, we have identified several genetic components of a novel biological pathway involved in the cellular transport of newly transcribed mRNAs and their subsequent local translocation. Importantly, in vivo biological validation has been achieved for two of these genes, pumilio and staufen. We propose to focus on this pathway via the following specific aims: 1. Molecular-genetic confirmation of selected genes in the pumilio/staufen pathway (staufen, oskar, moesin, pumilio, orb, elF2g, elFSc): We plan (i) genetic complementation with existing alleles and (ii) temporal and spatial rescue to establish a role for each of these genes in memory formation. 2. Protein expression studies of selected genes in the pumilio/staufen pathway: Immunostaining for most of these developmental genes never has been reported for the adult CNS. We will do so, with particular focus on subcellular co-localization experiments. 3. DNA microarray screen after cell purification: Studies focused on the neural substrates of olfactory memory continue to suggest the focus to be neurons of the mushroom bodies. Using a novel method, we plan to purify MB neurons from trained flies for DNA microarray experiments. We expect the signal from transcriptionally responsive genes to be greater in RNA isolated from this purified population of cells, thereby detecting additional neuronal participants of the pumilio/staufen pathway. Mechanisms of memory formation are remarkably conserved. Work in animal model systems can therefore greatly inform our understanding of human cognitive dysfunction. We will take advantage of the economy-of-scale and the molecular-genetic tools available in Drosophila to establish this novel biological pathway underlying memory formation. Our work in Drosophila will fuel studies of vertebrate homologs and eventually will contribute to our understanding of various forms of cognitive dysfunction in humans.

描述（由申请人提供）：LTM 形成的分子基础似乎在进化上是保守的。特别是，简单的学习形式似乎反映了神经元的保守细胞功能，而更复杂的行为任务则反映了跨物种神经元系统（解剖学）的多样性。尽管如此，这些复杂任务的记忆似乎激发了一种共同的潜在突触可塑性。因此，我们利用果蝇的规模经济和遗传工具的可用性，通过结合 DNA 微阵列和针对具有缺陷记忆的单基因突变体的行为筛选，重点发现参与 LTM 的下游基因。 ，我们已经确定了一种新的生物途径的几个遗传成分，该途径涉及新转录的 mRNA 的细胞运输及其随后的局部易位。重要的是，其中两个基因 pumilio 和 staufen 已经实现了体内生物学验证。 我们建议通过以下具体目标来关注这一途径： 1. pumilio/staufen途径中选定基因（staufen、oskar、moesin、pumilio、orb、elF2g、elFSc）的分子遗传学确认：我们计划（i）与现有等位基因的遗传互补和（ii）时间和空间救援确定每个基因在记忆形成中的作用。 2. pumilio/staufen 途径中选定基因的蛋白质表达研究：对于成人 CNS，大多数这些发育基因的免疫染色从未有过报道。我们将这样做，特别关注亚细胞共定位实验。 3.细胞纯化后的DNA微阵列筛选：针对嗅觉记忆的神经基质的研究继续表明焦点是蘑菇体的神经元。我们计划使用一种新方法从经过训练的果蝇中纯化 MB 神经元，用于 DNA 微阵列实验。我们预计从纯化的细胞群中分离出的 RNA 中来自转录反应基因的信号会更强，从而检测 pumilio/staufen 通路的其他神经元参与者。 记忆形成的机制非常保守。因此，动物模型系统的工作可以极大地促进我们对人类认知功能障碍的理解。我们将利用果蝇的规模经济和分子遗传学工具来建立这种记忆形成的新生物途径。我们在果蝇方面的工作将推动脊椎动物同源物的研究，并最终有助于我们了解人类各种形式的认知功能障碍。