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

影响果蝇伤害感受敏感性的犰狳/α-连环蛋白机制的研究

• 批准号: 10653377
• 负责人: GEOFFREY GANTER
• 金额: $ 42.6万
• 依托单位: UNIVERSITY OF NEW ENGLAND
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-08-01 至 2026-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10653377
• 关键词: Adaptor Signaling Protein; Address; Adherens Junction; Adhesions; Adhesives; Affect; Afferent Neurons; Analgesics; Behavior; Behavioral; Behavioral Assay; Biological Assay; Biological Models; Cadherins; Cell Adhesion; Cell-Cell Adhesion; Cells; Complement; Complex; Confocal Microscopy; Cytoskeleton; Dendrites; Drosophila genus; Emotional; Epidemic; Epidermis; Family Dasypodidae; Genes; Genetic; Genetic Models; Genetic Transcription; Health Promotion; Homologous Gene; Human; Hypersensitivity; Image Analysis; Impairment; Interneurons; Intractable Pain; Investigation; Larva; Life; Ligands; Mammals; Measures; Mechanics; Membrane; Methodology; Microscopy; Modeling; Neurons; Nociception; Nociceptors; Opiate Addiction; Opioid; Pain; Pain management; Pathway interactions; Pharmaceutical Preparations; Population; Positioning Attribute; Process; Proteins; Quality of life; RNA Interference; Reaction; Regulation; Rodent; Role; Secure; Sensory; Signal Pathway; Signal Transduction; Signaling Molecule; Source; Stimulus; Synapses; System; Testing; Tissues; Transcription Regulation Pathway; Transcriptional Regulation; Translations; Work; addiction liability; alpha catenin; arm; arm function; autocrine; beta catenin; candidate identification; clinical pain; economic impact; experimental arm; experimental study; fly; gene conservation; health warning; improved; knock-down; new therapeutic target; novel; novel therapeutics; overexpression; pain reduction; pain sensitivity; receptor; side effect; synaptogenesis; tool

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.

项目摘要 正常的疼痛通过警告我们潜在的组织损伤来促进健康，但不正常的疼痛会降低质量。 为全世界数以百万计的人提供生命的源泉。目前对疼痛的治疗是不够的，部分原因是 我们最好的止痛药阿片类药物的有害副作用。更好的治疗异常疼痛的方法是严重的 needed.我们建议通过利用强大的基因工具包， 果蝇模型果蝇幼虫对伤害性刺激的反应是利用一种逃避行为， 绝对不会错的螺旋卷这个模型系统已经被用来识别几十个组件， 伤害敏感性在果蝇中发现的许多信号成分与哺乳动物非常相似 同行初步结果表明，B-连环蛋白的苍蝇同系物，称为Armadillo（Arm）调节 伤害敏感性，但这种调节的机制目前尚不清楚。目标1将检验假设 Arm通过影响众所周知的Wnt/Wg转录控制来调节敏感性， 通路该测试将通过靶向RNAi沉默和/或过表达构建体来完成，所述RNAi沉默和/或过表达构建体是靶向RNAi沉默和/或过表达构建体的关键基因。 使用Gal 4/UAS系统特异性针对伤害感受器神经元的途径基因。任何由此产生的更改 将在聚焦于逃避的热和机械伤害感受测定中观察伤害感受敏感性 行为目的2将检验Arm和相关组分调节伤害性感受的独立假设。 通过Arm的细胞粘附功能提高敏感性。我们将使伤害感受器表达标记形式的Arm 和其他成分，可以通过免疫荧光显微镜分析，以研究神经元臂的 神经元-表皮粘附连接（称为鞘）和与CNS中间神经元的突触的贡献。 我们将过表达Arm和其他成分，并将鞘或突触中的任何变化 与伤害感受敏感性的关系。拟议的研究有可能 确定影响伤害感受敏感性的新机制和成分。由于高度的 果蝇和哺乳动物信号分子之间的功能保守性，通过这些鉴定的组分 这些实验可能代表治疗人类异常疼痛的新药物的目标。