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 正常疼痛通过警告我们潜在的组织损害来促进健康，但异常疼痛降低了质量 全世界数百万人的生活。目前可用的疼痛治疗不足，部分原因是 我们最好的镇痛药的有害副作用，阿片类药物。更好的异常疼痛治疗方法很严重 需要。我们建议通过利用强大的遗传工具包来揭示止痛药的新目标 果蝇模型。果蝇幼虫对有害刺激有反应 毫无疑问的开瓶器卷。该模型系统已用于识别数十个调节的组件 伤害性灵敏度。飞行中鉴定出的许多信号传导成分与哺乳动物非常相似 同行。初步结果表明，B-catenin的Fly同源物称为Armadillo（ARM）调节 伤害性敏感性，但目前尚不清楚该调节的机制。 AIM 1将检验假设 该手臂通过在众所周知的WNT/WG转录控制中发挥其影响来调节灵敏度 路径。该测试将通过靶向键的RNAi沉默和/或过表达构建体来完成 使用GAL4/UAS系统专门针对伤害感受器神经元的途径基因。任何结果更改 在关注逃生的热感受激活测定中将观察到伤害感受性的敏感性 行为。 AIM 2将检验独立假设，即ARM和相关组件调节伤害感受器 通过ARM的细胞粘附功能的灵敏度。我们将使伤害感受器表达标记的手臂形式 以及可以通过免疫荧光显微镜分析的其他成分来研究神经元组 对称为护套的神经元 - 表皮粘合剂的贡献和与CNS中间神经元突触的贡献。 我们将低于和过表达的臂和其他组件，并将鞘或突触的任何变化相关联 使用我们的行为测定法形成具有伤害感性的敏感性。拟议的研究有可能 确定影响伤害感受敏感性的新型机制和组件。由于高度 苍蝇和哺乳动物信号分子之间的功能保护，这些组件由这些鉴定 实验可能代表用于治疗人类异常疼痛的新型药物的靶标。