Project 2: Ganter

项目2：甘特

• 批准号: 8466106
• 负责人: GEOFFREY GANTER
• 金额: $ 26.93万
• 依托单位: UNIVERSITY OF NEW ENGLAND
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/8466106
• 关键词: Adult; Affect; Alleles; Animals; Area; Aversive Stimulus; Behavioral; Binding; Biological Assay; Biological Models; Cell Nucleus; Cells; Chemical Stimulation; Chemicals; Data; Dopamine; Drosophila genome; Drosophila genus; Drosophila melanogaster; Drug Addiction; Drug Delivery Systems; Drug abuse; Ecdysone; Elements; Eligibility Determination; Food; Future; Genes; Genetic Techniques; Genomics; Heating; Homologous Gene; Hormonal; Human; Hypersensitivity; Instruction; Knowledge; Larva; Lead; Learning; Libraries; Link; Mammals; Mechanics; Mediating; Membrane; Memory; Modality; Mutate; Mutation; Nature; Nervous system structure; Neurons; Nociception; Nuclear; Organism; Orthologous Gene; Pain; Pain management; Painless; Pathway interactions; Perception; Phenotype; Process; RNA Interference; Research Personnel; Role; Sensory; Sex Characteristics; Signal Transduction; Sleep; Steroids; Stimulus; Stress; Study models; System; TRP channel; Testing; Therapeutic; Variant; Vertebrates; Visual system structure; Work; chemical hypersensitivity; design; ecdysone receptor; fly; genome wide association study; genome-wide; human disease; interest; memory process; mustard oil; mutant; neurogenetics; non-genomic; novel; relating to nervous system; research study; response; steroid hormone; temperature sensitive mutant

The modulation of nociception by novel factors including the steroid hormone ecdysone in the fruit fly will be investigated. The fruit fly Drosophila melanogaster model system has produced a quantity of data relevant to higher functions of the human nervous system, including findings in memory processing, sleep, drug abuse and addiction. Exploration of the neurogenetic and hormonal mechanisms controlling nociception in flies will increase our understanding of pain in humans. Using a mutant allele of the ecdysoneless (ecd^) steroid availability gene, we have shown in preliminary studies that when ecdysone is depleted in adult flies, they become hypersensitive to noxious thermal and chemical stimulation. This suggests that steroid signaling has an antinociceptive role in the fly, as has been shown in mammals. This finding establishes the fruit fly as a valid model for the study of steroid modulation of nociception, with implications extending to higher organisms such as humans. We will now characterize the cellular and subcellular nature of ecdysone's role in nociception, and identify other novel genes that are connected to antinociception in the fly. Aim 1 will test for alterations in nociception in flies with manipulated ecdysone levels. A temperature sensitive mutant allele of ecdysoneless, along with other mutants, will be used to ask whether ecdysone levels affect perception of, and behavioral responses to, noxious thermal, chemical and mechanical stimuli. Aim 2 will examine the mode of nociceptive ecdysone signaling in ecdysone receptor mutants. Experiments are designed to test whether disruption of ecdysone signaling at the level of the nucleus or at the membrane affects nociception, thereby determining if the mechanism of nociceptive modulation is a genomic or membrane process. Aim 3 will test for steroid modulation of identified nociceptive neurons. The potential role of ecdysone in the activity of known TRP channel-expressing nociceptive neurons will be assessed by temporal- and cell-specific blockade of signaling using genetic techniques. A parallel electrophysiological approach will also be used to analyze the response of TRP channel-expressing nociceptive neurons to varying levels of the steroid ecdysone. Aim 4 will screen a genome-wide mutant library to identify novel genes connected to antinociception in the fly. We will use a simple food-choice assay to rapidly identify mutants that are hypersensitive to a noxious chemical stimulus (mustard oil). This work should point to potentially novel human therapeutic drug targets.

通过新的因素调节伤害感受，包括果蝇中的类固醇激素ecdysone 调查。果蝇果蝇Melanogaster模型系统产生了一定数量的数据 人类神经系统的较高功能，包括记忆加工，睡眠，滥用药物的发现 和成瘾。探索控制苍蝇伤伤心吸引力的神经遗传学和激素机制 提高我们对人类疼痛的理解。使用ecdysoneless（ECD^）类固醇的突变等位基因 可用性基因，我们在初步研究中表明，当埃dy虫在成年苍蝇中耗尽时，它们 对有害的热和化学刺激过敏。这表明类固醇信号传导 如哺乳动物所示，在苍蝇中具有抗伤害感受的作用。这一发现确立了果蝇 研究类固醇调节伤害感受的有效模型，含义扩展到更高 人类等有机体。我们现在将表征ecdysone角色的细胞和亚细胞性质 在伤害感受中，并识别其他与触觉抗伤害感受相关的新型基因。 AIM 1将测试 为了改变操纵cdysone水平的苍蝇伤害感受。温度敏感的突变等位基因 ecdysoneless和其他突变体将被用来询问ecdysone水平是否影响对的感知 以及对有害热，化学和机械刺激的行为反应。 AIM 2将检查 ecdysone受体突变体中的伤害感受ecdysone信号传导的模式。实验旨在测试 ecdysone信号在核水平上的破坏还是在膜上影响伤害感受， 从而确定伤害性调节的机制是基因组或膜过程。目标3 将测试确定的伤害性神经元的类固醇调节。 ecdysone在活动中的潜在作用 将通过时间和细胞特异性评估已知TRP通道的伤害性神经元 使用遗传技术阻断信号传导。平行电生理方法也将用于 分析表达TRP通道的伤害性神经元对类固醇水平的反应 ecdysone。 AIM 4将筛选全基因组突变库，以鉴定与 触觉中的抗伤害感受。我们将使用简单的食品选择测定法来快速识别突变体 对有害化学刺激（芥末油）的高度敏感。这项工作应该指向潜在的新颖 人类治疗药物靶标。