Project 2: Ganter

项目2：甘特

• 批准号: 8529581
• 负责人: GEOFFREY GANTER
• 金额: $ 20.95万
• 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/8529581
• 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 Targeting; 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.

包括果蝇中的类固醇激素蜕皮激素在内的新因子对伤害感受的调节将是 研究了果蝇Drosophila melanogaster模型系统已经产生了大量的数据， 人类神经系统的高级功能，包括记忆处理，睡眠，药物滥用 和上瘾。对控制果蝇伤害感受的神经发生和激素机制的探索， 增加我们对人类疼痛的理解。使用无蜕皮激素（ecd^）类固醇的突变等位基因 利用率基因，我们在初步研究中表明，当蜕皮激素在成年苍蝇中耗尽时， 对有害的热和化学刺激变得高度敏感。这表明类固醇信号 在果蝇中具有抗伤害感受作用，如在哺乳动物中所示。这一发现确立了果蝇作为 一个有效的模型，研究类固醇调制的伤害性感受，其影响延伸到更高 生物，如人类。我们现在将描述蜕皮激素作用的细胞和亚细胞性质 在伤害感受，并确定其他新的基因是连接到抗伤害感受的苍蝇。目标1将测试 用于改变具有操纵的蜕皮激素水平的果蝇的伤害感受。一个温度敏感突变等位基因 无蜕皮激素的，沿着其他突变体，将被用来询问蜕皮激素水平是否影响对， 以及对有害的热、化学和机械刺激的行为反应。目标2将审查 蜕皮激素受体突变体中的伤害性蜕皮激素信号传导模式。实验旨在测试 无论在细胞核水平还是在细胞膜水平上破坏蜕皮激素信号传导影响伤害感受， 从而确定伤害感受调节的机制是基因组过程还是膜过程。目标3 将测试类固醇对已鉴定的伤害感受神经元的调节。蜕皮激素在活性中的潜在作用 已知的TRP通道表达伤害性神经元将通过时间和细胞特异性的 利用遗传技术阻断信号传导。还将使用平行电生理方法， 分析TRP通道表达伤害感受神经元对不同水平类固醇的反应 蜕皮激素Aim 4将筛选全基因组突变体文库，以鉴定与 果蝇的抗伤害感受我们将使用一个简单的食物选择试验来快速识别突变体， 对有害的化学刺激（芥子油）过敏。这项工作应该指出潜在的新颖性 人治疗药物靶点。