Role of glia in ethanol tolerance in Drosophila

胶质细胞在果蝇乙醇耐受中的作用

• 批准号: 8100512
• 负责人: FREDERICK W WOLF
• 金额: $ 19.26万
• 依托单位: ERNEST GALLO CLINIC AND RESEARCH CENTER
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-30 至 2011-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8100512
• 关键词: A kinase anchoring protein; Acute; Adult; Affect; Animal Model; Attention; Behavioral; Blood - brain barrier anatomy; Brain; Cell physiology; Cells; Communication; Complex; Coupled; Cyclic AMP-Dependent Protein Kinases; Cytoskeleton; Defect; Development; Dose; Drosophila genus; Ethanol; Exhibits; Exposure to; GTP-Binding Proteins; Gene Expression Regulation; Genes; Genetic; Genetic Screening; Goals; Human; Learning; Link; Mammals; Mediating; Methodology; Methods; Molecular; Molecular Target; Mus; Mutation; Nervous System Physiology; Neuraxis; Neuroglia; Neurons; Neurophysiology - biologic function; Organism; Phenotype; Physiological; Play; Property; Protein Kinase C; Proteins; Reagent; Research; Role; Scaffolding Protein; Shapes; Signal Pathway; Signal Transduction; Signal Transduction Pathway; Signaling Molecule; Structure; System; Tertiary Protein Structure; Time; Vertebrates; alcohol effect; alcohol exposure; alcohol response; alcohol sensitivity; alcohol use disorder; base; fly; gene function; genetic manipulation; insight; intercellular communication; loss of function; molecular marker; mutant; neural circuit; novel; public health relevance; response; tool

DESCRIPTION (provided by applicant): Glial cells are crucial for the normal functioning of the brain, communicate extensively with neurons, and are affected by ethanol exposure, yet their role in the development of alcohol use disorders is poorly understood. The goal of this proposal is to study the contribution of glia to ethanol behavioral responses in the fruit fly Drosophila. A microarray study of gene regulation by ethanol exposure coupled with a genetic screen of flies carrying mutations in ethanol-responsive genes led to the identification of novel genes required for ethanol tolerance. Preliminary studies of one of these, the A-kinase anchoring protein Akap200, indicate that it is exclusively localized to glia in adult flies and that its function is required in glia to promote the development of ethanol tolerance. AKAP proteins spatially and temporally coordinate protein kinase A signaling with other cell signaling pathways, including protein kinase C, and we found that both protein kinase A and protein kinase C, like Akap200, can promote ethanol tolerance in glia. Additionally, we found that the glial blood-brain barrier, where Akap200 is prominently expressed, responds to ethanol exposure by changing its morphological and functional properties, and that barrier function is partially compromised in Akap200 mutants. We propose that glia regulate ethanol tolerance through conserved signaling pathways that are coordinated by Akap200. There are three main goals of our proposed research. First, we plan to characterize the response of each class of central nervous system glia to ethanol exposure by determining the time and dose-dependent effects of ethanol on glial structure/function, and the role played by glial cellular processes in the development of ethanol behavioral responses. Second, we plan to characterize the role of Akap200 and Akap200-interacting proteins in the development of ethanol tolerance and glial structure/function in order to establish the mechanisms by which glia regulate ethanol behavioral responses. Third, we plan to carry out a genetic screen for novel glial genes that regulate ethanol behavioral responses. The recent availability of tools for genetically targeting specific glial types, for limiting genetic manipulations to adults, and methodology for assessing glial structure and function, coupled with our discovery of a glial protein kinase A signaling pathway for ethanol tolerance, has given us an opportunity to make rapid progress in understanding the contributions of glia to ethanol behavioral responses. Evolutionary conservation of protein kinase A signaling and many glial properties suggests that what is learned in Drosophila can transfer to mammalian systems and may impact the treatment of alcohol use disorders in humans. PUBLIC HEALTH RELEVANCE: Glial cells are critically important for the normal functioning of the brain and they show complex and dynamic responses to ethanol, but their role in the development of alcohol use disorders remains to be characterized. We found that glia in the fruit fly Drosophila alter their shape and function in response to ethanol, and that evolutionarily conserved signal transduction pathways act in fly glia to regulate ethanol tolerance. Sophisticated genetic tools, a high level of molecular conservation, and similar glial properties make Drosophila an ideal system for uncovering molecular mechanisms that can provide novel insights in vertebrates, including humans.

描述（由申请人提供）：神经胶质细胞对大脑的正常功能至关重要，与神经元广泛沟通，并受到乙醇暴露的影响，但其在酒精使用障碍发展中的作用知之甚少。这个提议的目的是研究神经胶质细胞对果蝇乙醇行为反应的贡献。乙醇暴露的基因调控的微阵列研究加上携带突变的果蝇的乙醇反应基因的遗传筛选，导致乙醇耐受性所需的新基因的鉴定。其中之一，A-激酶锚定蛋白Akap 200的初步研究表明，它是专门本地化的神经胶质在成年苍蝇，它的功能是必需的神经胶质细胞，以促进乙醇耐受性的发展。AKAP蛋白在空间和时间上协调蛋白激酶A信号传导与其他细胞信号传导途径，包括蛋白激酶C，我们发现蛋白激酶A和蛋白激酶C，如Akap 200，可以促进神经胶质细胞的乙醇耐受性。此外，我们发现，神经胶质血脑屏障，其中Akap 200的显着表达，响应乙醇暴露通过改变其形态和功能特性，和屏障功能是部分受损的Akap 200突变体。我们认为神经胶质细胞通过Akap 200协调的保守信号通路调节乙醇耐受性。我们提出的研究有三个主要目标。首先，我们计划通过确定乙醇对神经胶质结构/功能的时间和剂量依赖性影响，以及神经胶质细胞过程在乙醇行为反应发展中所起的作用，来表征每一类中枢神经系统神经胶质细胞对乙醇暴露的反应。其次，我们计划描述Akap 200和Akap 200相互作用蛋白在乙醇耐受性和神经胶质结构/功能发展中的作用，以建立神经胶质调节乙醇行为反应的机制。第三，我们计划对调节乙醇行为反应的新型神经胶质基因进行遗传筛选。最近可用的工具，遗传靶向特定的神经胶质细胞类型，限制遗传操作的成年人，和方法评估神经胶质细胞的结构和功能，再加上我们发现的神经胶质蛋白激酶A信号通路的乙醇耐受性，给了我们一个机会，使快速进展，了解神经胶质细胞乙醇行为反应的贡献。蛋白激酶A信号和许多神经胶质特性的进化保守性表明，在果蝇中学到的东西可以转移到哺乳动物系统，并可能影响人类酒精使用障碍的治疗。 公共卫生相关性：神经胶质细胞对大脑的正常功能至关重要，它们对乙醇表现出复杂和动态的反应，但它们在酒精使用障碍发展中的作用仍有待表征。我们发现果蝇中的神经胶质细胞改变了它们的形状和功能，以响应乙醇，并且进化上保守的信号转导途径在果蝇神经胶质细胞中起作用以调节乙醇耐受性。复杂的遗传工具，高水平的分子保守性和类似的神经胶质特性使果蝇成为揭示分子机制的理想系统，可以为脊椎动物（包括人类）提供新的见解。