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.

包括类固醇激素蜕皮激素在内的新因素对果蝇伤害感受的调节将是 调查过了。果蝇黑腹果蝇模型系统已经产生了大量与 人类神经系统的高级功能，包括记忆处理、睡眠、药物滥用 还有毒瘾。果蝇意志中控制伤害性感受的神经遗传学和激素机制探讨 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 可利用性基因，我们在初步研究中表明，当成年果蝇的蜕皮激素耗尽时，它们 对有害的热刺激和化学刺激过敏。这表明类固醇信号 在果蝇中具有抗伤害作用，正如在哺乳动物中所显示的那样。这一发现证实了果蝇是 研究类固醇对伤害性感觉的调制的有效模型，其意义延伸到更高 生物体，如人类。我们现在将描述蜕皮激素的细胞和亚细胞性质 并找出其他与果蝇抗伤害感受相关的新基因。AIM 1将测试 用于改变被操纵的蜕皮激素水平的果蝇的伤害感受。一种温度敏感突变等位基因 与其他突变体一起，将被用来询问蜕皮激素水平是否会影响感知， 以及对有害的热、化学和机械刺激的行为反应。Aim 2将研究 蜕皮激素受体突变体的伤害性蜕皮激素信号转导方式。实验是为了测试 蜕皮激素信号在核水平或膜水平的中断是否会影响伤害性感受， 从而确定伤害性调制的机制是基因组过程还是膜过程。目标3 将测试类固醇对已识别的伤害性神经元的调制。蜕皮激素在活动中的潜在作用 已知的表达Trp通道的伤害性神经元将通过特定的时间和细胞进行评估 利用基因技术阻止信号传递。一种并行的电生理学方法也将用于 分析表达Trp通道的伤害性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文库，以确定与 苍蝇体内的抗伤害感受。我们将使用一种简单的食物选择试验来快速鉴定 对有毒的化学刺激(芥子油)过敏。这项工作应该指向潜在的新奇 人类治疗性药物靶点。