Investigation of Armadillo/ß-catenin Mechanisms Influencing Nociceptive Sensitivity in Drosophila

项目摘要 正常疼痛通过警告我们潜在的组织损伤来促进健康，但异常疼痛会降低质量。 世界各地数百万人的生活。目前对疼痛的可用治疗是不够的，部分原因是 我们最好的止痛药阿片类药物的有害副作用。对于异常疼痛的更好的治疗方法是糟糕的 需要的。我们建议通过开发强大的基因工具包来揭示止痛药的新靶点 果蝇模型。果蝇幼虫对有害刺激的反应是通过一种逃逸行为，这种行为包括 无可辩驳的开瓶器滚筒。这个模型系统已经被用来识别数十个调节 伤害性敏感。在苍蝇体内发现的许多信号成分与它们的哺乳动物非常相似 对口单位。初步结果表明，果蝇中B-连环蛋白的同系物，称为Armadillo(ARM)，调节 伤害性敏感，但这一调节机制目前尚不清楚。目标1将检验这一假设 该臂通过在众所周知的Wnt/Wg转录调控中发挥影响来调节敏感性 路径。这项测试将通过靶向RNAi沉默和/或Key的过度表达构造来完成 利用Gal4/UAS系统对伤害性感受器神经元特异的通路基因。任何由此产生的更改 伤害性敏感性将在关注逃逸的热和机械伤害性测试中观察到。 行为。目标2将检验ARM及其相关成分调节伤害性感受的独立假说 通过ARM的细胞黏附功能实现敏感性。我们将使伤害性感受器表达标记形式的手臂 以及其他可以通过免疫荧光显微镜分析来研究神经臂的成分 对称为鞘的神经元-表皮粘连连接和与中枢神经系统中间神经元的突触的贡献。 我们将过低和过表达ARM和其他组件，并将鞘或突触的任何变化关联起来 使用我们的行为分析形成伤害性敏感性。拟议的研究有可能 确定影响伤害性敏感性的新机制和成分。因为高度的 苍蝇和哺乳动物信号分子之间的功能保守，由这些识别的成分 实验可能代表着治疗人类异常疼痛的新药的靶点。

Project Summary Normal pain promotes health by warning us of potential tissue damage, but abnormal pain reduces the quality of life for millions around the world. Available treatment for pain is currently inadequate, in part because of the deleterious side effects of our best analgesics, the opioids. Better treatments for abnormal pain are badly needed. We propose to reveal novel targets for pain medications by exploiting the powerful genetic toolkit of the Drosophila model. Fruit fly larvae react to noxious stimuli using an escape behavior consisting of an unmistakable corkscrew roll. This model system has been used to identify dozens of components that regulate nociceptive sensitivity. Many signaling components identified in the fly are very similar to their mammalian counterparts. Preliminary results indicate that the fly homolog of B-catenin, called Armadillo (Arm) regulates nociceptive sensitivity but the mechanism of this regulation is currently unclear. Aim 1 will test the hypothesis that Arm regulates sensitivity by exerting its influence in the well-known Wnt/Wg transcriptional control pathway. This testing will be accomplished by targeting RNAi silencing and/or overexpression constructs of key pathway genes specifically to the nociceptor neurons using the Gal4/UAS system. Any resulting changes to nociceptive sensitivity will be observed in thermo- and mechanonociception assays that focus on the escape behavior. Aim 2 will test the independent hypothesis that Arm and related components regulate nociceptive sensitivity through Arm's cell adhesion function. We will cause the nociceptors to express tagged forms of Arm and other components that can be analyzed by immunofluorescent microscopy to study neuronal Arm's contribution to neuronal-epidermal adhesive junctions called sheaths and to synapses with CNS interneurons. We will under- and overexpress Arm and other components and correlate any changes in sheath or synapse formation with nociceptive sensitivity using our behavioral assays. The proposed studies have the potential to identify novel mechanisms and components that affect nociceptive sensitivity. Because of the high degree of functional conservation between fly and mammalian signaling molecules, components identified by these experiments may represent targets for novel medications for the treatment of abnormal pain in humans.