GPCR-mediated nongenomic steroid actions in control of alcohol-induced behavior

GPCR 介导的非基因组类固醇作用控制酒精诱发的行为

• 批准号: 8396210
• 负责人: Emily Petruccelli
• 金额: $ 2.85万
• 依托单位: UNIVERSITY OF IOWA
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-07-01 至 2015-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8396210
• 关键词: Address; Adrenergic Receptor; Affect; Animal Model; Animals; Antibodies; Behavior; Behavior Control; Behavioral; Binding; Biological; Brain; Catecholamines; Cell physiology; Complementary DNA; Cyclic AMP; Development; Disease; Dopamine; Drosophila genus; Drosophila melanogaster; Ecdysone; Equilibrium; Ethanol; Exposure to; Future; G-Protein-Coupled Receptors; Genetic; Genetic Markers; Genetic Transcription; Genomics; Goals; Insecta; Knowledge; Learning; MAP Kinase Gene; Mediating; Mission; Molecular; Nervous System Physiology; Nervous system structure; Neurobiology; Neurologic; Neurons; Nuclear Hormone Receptors; Nuclear Receptors; Outcome; Outcome Study; Pathway interactions; Pattern; Pharmacology; Phenocopy; Phenotype; Physiological; Play; Posture; Prevention; Prevention strategy; RNA Interference; Regulation; Research; Role; Sedation procedure; Signal Pathway; Signal Transduction; Site; Steroid Receptors; Steroids; System; Techniques; Testing; Work; alcohol behavior; alcohol exposure; alcohol response; alcohol use disorder; analytical method; base; experience; fly; genetic variant; in vivo; innovation; insight; knowledge base; mutant; neurogenetics; neuropsychiatry; non-genomic; novel; post-doctoral training; receptor; research study; steroid hormone; tool; transcription factor

DESCRIPTION (provided by applicant): Steroid hormones have significant impact on the development and function of the nervous system and play crucial roles in various neurological and neuropsychiatric disorders. While steroids primarily act through the "genomic" mechanism by binding nuclear hormone receptors and activating transcription, accumulated evidence indicates that steroid hormones also induce biological effects rapidly and independent of their nuclear receptors. Despite the potential importance of such "nongenomic" steroid actions, their functional significance and underlying molecular mechanisms are still poorly understood. To address this gap in our knowledge, DopEcR, a recently discovered G-protein coupled receptor (GPCR), will be studied using genetic tools uniquely available in the fruit fly in Drosophila melanogaster. DopEcR is a strong candidate for a nonclassical steroid receptor, because this GPCR can directly bind and respond to the insect steroid hormone ecdysone as well as the catecholamine dopamine. Although DopEcR is known to have the highest sequence similarity with vertebrate ¿-adrenergic receptors and is preferentially expressed in the nervous system, its function in intact animals is not known. The objective of this proposed research is to reveal the roles of DopEcR in behavioral regulation and identify its underlying mechanism of actions. The central hypothesis of this proposed research is that DopEcR-mediated ecdysone signaling plays a role in controlling balance of excitation and inhibition upon exposure to ethanol by modulating the effects of dopamine, cAMP and MAPK pathways. This hypothesis is based on my recent discovery that newly identified DopEcR mutants display intriguing phenotypes in ethanol-induced behaviors. The two specific aims that will be pursued in this project are: 1) Determine the endogenous expression patterns and functional sites of action of DopEcR, and 2) Determine the mechanisms by which DopEcR regulates behavioral responses to ethanol. For Aim 1, behavioral analyses will be carried out following the expression of wild type cDNA or RNAi for DopEcR in a neuronal subset-specific manner. For Aim 2, genetic variants of DopEcR will be examined for their interactions with dopamine, cAMP and MAPK pathways. Successful completion of these aims is expected to reveal how GPCR-mediated nongenomic steroid signaling controls ethanol-induced behavior in Drosophila. The outcomes of this application will have an important positive impact because they are expected to provide strong mechanism-based in vivo evidence for the nongenomic effects of steroid hormones on behavioral regulation. Due to the high functional and mechanistic conservation of steroid signaling and alcohol response behaviors, understanding these effects should provide novel insights into the future prevention and treatment of alcohol use disorders. PUBLIC HEALTH RELEVANCE: The proposed studies aim to understand the underappreciated molecular and cellular processes by which steroid hormones control behavioral responses to alcohol. This project is highly relevant to NIH's mission because the findings are expected to ultimately contribute toward the development of innovative strategies for the prevention and treatment of alcohol use disorders as well as other diseases that are significantly affected by steroids.

描述（由申请人提供）：类固醇激素对神经系统的发育和功能有显著影响，并在各种神经系统和神经精神疾病中发挥关键作用。虽然类固醇主要通过结合核激素受体和激活转录的“基因组”机制起作用，但积累的证据表明，类固醇激素也迅速诱导生物效应，且不依赖于其核受体。尽管这种“非基因组”类固醇作用的潜在重要性，其功能意义和潜在的分子机制仍然知之甚少。为了解决我们知识中的这一空白，DopEcR，最近发现的G蛋白偶联受体（GPCR），将使用遗传工具在果蝇中唯一可用的果蝇进行研究。DopEcR是非经典类固醇受体的强有力的候选者，因为该GPCR可以直接结合并响应昆虫类固醇激素蜕皮激素以及儿茶酚胺多巴胺。虽然已知DopEcR与脊椎动物的肾上腺素能受体具有最高的序列相似性，并且优先在神经系统中表达，但其在完整动物中的功能尚不清楚。这项研究的目的是揭示DopEcR在行为调节中的作用，并确定其潜在的作用机制。这项研究的中心假设是DopEcR介导的蜕皮激素信号在控制暴露于乙醇后的兴奋和抑制平衡中发挥作用，通过调节多巴胺，cAMP和MAPK通路的作用。这一假设是基于我最近的发现，即新发现的DopEcR突变体在乙醇诱导的行为中显示出有趣的表型。本项目将追求的两个具体目标是：1）确定DopEcR的内源性表达模式和作用的功能位点，以及2）确定DopEcR调节乙醇行为反应的机制。对于目标1，将在以神经元亚群特异性方式表达DopEcR的野生型cDNA或RNAi后进行行为分析。对于目标2，将检查DopEcR的遗传变体与多巴胺、cAMP和MAPK途径的相互作用。这些目标的成功完成有望揭示GPCR介导的非基因组类固醇信号传导如何控制乙醇诱导的果蝇行为。该应用的结果将产生重要的积极影响，因为它们有望为类固醇激素对行为调节的非基因组效应提供强有力的基于机制的体内证据。由于类固醇信号传导和酒精反应行为的高功能和机械保守性，了解这些影响应该为未来预防和治疗酒精使用障碍提供新的见解。 公共卫生相关性：这些研究旨在了解类固醇激素控制酒精行为反应的分子和细胞过程。该项目与NIH的使命高度相关，因为这些发现有望最终有助于制定预防和治疗酒精使用障碍以及其他受类固醇显著影响的疾病的创新策略。