GENOME RESPONSES TO DROSOPHILA ADDICTION USING DNA CHIPS

使用 DNA 芯片研究果蝇成瘾的基因组反应

• 批准号: 6291515
• 负责人: TIM P TULLY
• 金额: $ 56.73万
• 依托单位: COLD SPRING HARBOR LABORATORY
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-09-28 至 2003-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6291515
• 关键词: DNA; Drosophilidae; alcoholism /alcohol abuse; behavioral /social science research tag; behavioral genetics; cocaine; cyclic AMP; drug addiction; ethanol; ethology; gene mutation; genome; in situ hybridization; laboratory rabbit; microarray technology; northern blottings; pharmacogenetics; polymerase chain reaction

DESCRIPTION: (Applicant's Abstract) Mechanisms of drug addiction appear to be remarkably conserved. Work in animal model systems can therefore greatly inform our understanding of human addiction. The economy-of-scale and molecular-genetic tools available in fruit flies will be used to identify genes and genetic pathways of addiction. Recent studies in Drosophila have established behavioral sensitivity models for both cocaine and ethanol. In the latter, an allele of amnesiac, a learning and memory gene that has some homology to a vertebrate neuropeptide that stimulates cAMP signaling, was identified. Amnesiac mutants show decreased tolerance to ethanol, a defect that can be reversed with pharmacological stimulation of cAMP levels. In the former, several single-gene mutants were shown to affect cocaine sensitization, which also was mediated by dopaminergic signaling. These studies validate Drosophila as a genetic model system for both cocaine and alcohol addiction and implicate the cAMP signaling pathway in the response to both. This Drosophila model system of addiction will be enhanced with the use of DNA microarray chips to identify drug-related changes in gene expression on a genomic-wide scale. In preliminary experiments, a partial genome prototype Drosophila chip has been used to identify 176 candidate genes transcriptionally altered in amnesiac mutants and 75 that are transcriptionally altered by pharmacological manipulation of cAMP levels. We have confirmed some of the statistical candidates from the chip using either Northern blot analysis or quantitative PCR. From initial analyses of more than 100 chips, we also have developed a novel and general statistical method to analyze DNA chip data. These initial results now will be integrated with data from exposures to cocaine and ethanol. Central to our approach is the comparison of three genomic responses: (1) drug-exposed vs. control, (2) pharmacological manipulation vs. control and (3) mutant vs. wild-type. Chosen first for further in vivo study will be those genes that are transcriptionally responsive in all three contexts - and to both cocaine and ethanol exposures. Given the "homology of function" already observed for drug responses in flies and vertebrates, the genes identified in flies most likely will have homologues in vertebrates. Hence this work will lead directly to work in mammalian models of addiction and ultimately will yield a greater understanding of the genetic basis of addiction in humans.

描述：（申请人的摘要）药物成瘾的机制似乎是 非常保守。因此，在动物模型系统中的工作可以极大地告知 我们对人类成瘾的理解。尺度和 水果蝇中可用的分子遗传工具将用于识别基因 和成瘾的遗传途径。果蝇最近的研究 为可卡因和乙醇建立了行为敏感性模型。在 后者是失忆症的等位基因，一种学习和记忆基因，有一些 与刺激cAMP信号传导的脊椎动物神经肽的同源性是 确定。失忆突变体显示对乙醇的耐受性降低，这是一个缺陷 可以通过对营地水平的药理学刺激来逆转。在前者中 显示几种单基因突变体会影响可卡因敏化，这 还通过多巴胺能信号传导介导。这些研究验证了果蝇 作为可卡因和酒精成瘾的遗传模型系统，并暗示 对两者的响应中的营地信号通路。 使用DNA将增强这种果蝇模型的成瘾模型 微阵列芯片以鉴定在A上与药物相关的基因表达变化 基因组范围。在初步实验中，部分基因组原型 果蝇芯片已用于鉴定176个候选基因转录 在失忆突变体和75种因转录改变而改变的改变 营地水平的药理学操纵。我们已经确认了一些 使用Northern印迹分析或 定量PCR。从最初的100多个芯片分析，我们也有 开发了一种新颖的一般统计方法来分析DNA芯片数据。 这些初始结果现在将与从暴露到的数据集成到 可卡因和乙醇。我们方法的核心是三个基因组的比较 反应：（1）暴露于药物与对照，（2）药理学操纵vs。 控制和（3）突变体与野生型。首先选择进一步的体内研究 将是那些在所有三种情况下转录响应敏感的基因 - 以及可卡因和乙醇暴露。鉴于“功能同源” 已经观察到蝇和脊椎动物中的药物反应，这些基因 在苍蝇中识别的很可能会在脊椎动物中具有同源物。因此 工作将直接导致在成瘾的哺乳动物模型中工作 将对人类成瘾的遗传基础有更深入的了解。