Regulation of Learning and Memory by Immediate Early Genes in Drosophila brain

描述(申请人提供)：虽然学习和记忆背后的几种细胞机制已经被很好地研究了，但对有助于正确记忆形成、保持和提取的遗传成分知之甚少。一类特殊的基因被称为即刻早期基因(IEGs)，它在神经元中快速表达，对活动做出反应，并与学习和记忆有关。我们已经开始了对果蝇大脑的全基因组分析，以发现可能构成记忆基础的新的IEG。我们的初步结果显示，在果蝇的大脑中，有278个候选IEG在感受刺激后有不同的表达，到目前为止，我们已经使用高通量的行为筛查，发现其中28个基因与学习和记忆有关。除了经典的IEG对神经元活动的反应迅速上调外，我们还识别了87个在刺激后立即下调的基因，其中几个基因与我们的行为屏幕上的记忆有关。这项提议的目标是描绘我们识别的6个在人类中保守的新IEG对学习和记忆所必需的分子机制。我们选择了3个上调的基因(CG11964、Sallimus和淀粉酶远端)和3个下调的基因(热休克蛋白-70Ab、热休克蛋白-70BA和Maggie)，它们具有不同的分子功能，这将为理解记忆形成和恢复所需的不同分子和细胞机制提供基础。我们将使用标准的果蝇工具和技术，如替代等位基因系和缺陷库，验证我们的IEG对于学习至关重要，我们将使用RNA探针和抗体绘制基因在大脑中的表达图，以描绘几种不同类型的学习所需的特定区域。我们还将通过在发育过程中用RNAi抑制基因表达来研究这些基因的潜在发育作用。我们的研究结果将具体说明这6个特定的新基因对正常记忆功能至关重要的方法。我们的结果还将提供对正常学习和记忆机制的更多了解，以及导致记忆丧失的神经疾病的潜在治疗目标。

DESCRIPTION (provided by applicant): While several cellular mechanisms that underlie learning and memory have been well-studied, little is known about the genetic components that contribute to proper memory formation, retention, and retrieval. A special class of genes called immediate early genes (IEGs) is rapidly expressed in neurons in response to activity, and has been implicated in learning and memory. We have initiated a genome-wide analysis of Drosophila melanogaster brains in order to discover novel IEGs that may underlie memory. Our preliminary results have revealed 278 candidate IEGs that are expressed differently in the Drosophila brain after exposure to sensory stimulation, and we have used a high-throughput behavior screen to implicate 28 of these genes so far in learning and memory. In addition to the classical IEGs that are rapidly up-regulated in response to neuronal activity, we have also identified 87 genes that are down-regulated immediately following stimulation, several of which we have implicated in memory using our behavior screen. The goal of this proposal is to delineate the molecular mechanisms by which 6 novel IEGs that we identified, that are conserved in humans, are necessary for learning and memory. We have selected 3 up-regulated genes (CG11964, sallimus, and Amylase distal) and 3 down-regulated genes (Heat-shock-protein-70Ab, Heat-shock-protein- 70Ba, and maggie) that have a variety of molecular functions, which should provide an understanding of the diverse molecular and cellular mechanisms necessary for memory formation and retrieval. We will validate that our IEGs are critical for learning using standard Drosophila tools and techniques such as alternative allele lines and deficiency stocks, and we will map expression of the genes in the brain using RNA probes and antibodies to delineate specific regions that are needed for several different types of learning. We will also investigate a potential developmental role for the genes by knocking down gene expression with RNAi following development. The outcome of our studies will specify the methods by which these 6 specific novel genes are critical for proper memory function. Our results will also provide a greater understanding of normal learning and memory mechanisms, as well as potential therapeutic targets for neurological diseases that result in memory loss.