A Genome-Wide Analysis of Nociception Molecules, from Expression to Function

描述(由申请人提供)：彻底了解伤害性感觉神经元的分子机制将有助于更好地理解急性疼痛的保护性分子机制。为了识别伤害性分子，我们之前使用果蝇黑腹果蝇进行了伤害性缺陷突变体的遗传筛选。这导致我们发现了无痛基因，它在伤害性感觉神经元中显示出丰富的表达，是急性伤害性感受所必需的。这种无痛基因以及一种名为dTRPA1的相关基因编码TRPA1的同源物。人类TRPA1基因突变导致家族性阵发性疼痛综合征(FEPS)，并且TRPA1在伤害性感受器神经元中高表达。我们发现扒手是机械伤害性感受的关键调节因子，而机械伤害性感受在果蝇的伤害性感受器中特异表达。本研究的目的是鉴定果蝇伤害性感受器神经元中表达的其他基因，并测试这些基因在伤害性感受中的功能需求。根据我们最近的研究，我们确定IV类多树突状神经元是果蝇幼虫的多模式伤害性感受器，我们将：1)确定伤害性IV类多树突状神经元的完整转录组。2)在体内利用组织特异性RNAi敲除伤害性感觉神经元中丰富表达的基因来检测这些细胞的功能需求。3.)从功能上描述我们在AIMS 1和AIMS 2中发现的对伤害性感受至关重要的新基因的子集。短期内，这些研究将使用高通量的方法对伤害性感受的分子和细胞机制提供关键的洞察，该方法是利用果蝇实现的。从长远来看，这些研究可能最终允许识别对人类伤害性感觉有贡献的进化保守机制。好了！

DESCRIPTION (provided by applicant): A thorough understanding of the molecular machinery that operates in nociceptive sensory neurons will lead to greater understanding of the protective molecular mechanisms of acute pain. To identify nociception molecules, we previously carried out a genetic screen for nociception defective mutants using the fruitfly Drosophila melanogaster. This led to our discovery of the painless gene, which shows enriched expression in nociceptive sensory neurons and is required for acute nociception. The painless gene, as well as a related gene named dTRPA1, encode homologues of TRPA1. Human mutations in TRPA1 mutations lead to familial episodic pain syndrome (FEPS) and TRPA1 is highly expressed in nociceptor neurons. We have identified pickpocket as a critical regulator of mechanical nociception which is specifically expressed in Drosophila nociceptors. The purpose of this proposal is to identify other genes expressed in nociceptor neurons of Drosophila and to test the functional requirement of those genes in nociception. Building upon our recent studies that identified the Class IV multidendritic neurons as polymodal nociceptors of the Drosophila larva we will: 1) Determine the complete transcriptome of nociceptive Class IV multidendritic neurons. 2) Test the functional requirement of genes with enriched expression in nociceptive sensory neurons using tissue specific RNAi knockdown in these cells in vivo. 3.) Functionally characterize a subset of novel genes that we identify to be essential for nociception in Aims 1 and 2. In the short term, these studies will provide critical insight into molecular and cellular mechanisms of nociception using a high throughput approach that is made possible using Drosophila. In the long term, these studies may eventually allow for the identification of evolutionarily conserved mechanisms that contribute to human nociception. !