Regulation of D1 Dopamine Receptor Expression by ncRNA in Cocaine Addiction

可卡因成瘾中 ncRNA 对 D1 多巴胺受体表达的调节

• 批准号: 7776977
• 负责人: ELDO V KUZHIKANDATHIL
• 金额: $ 14.81万
• 依托单位: UNIV OF MED/DENT OF NJ-NJ MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-03-01 至 2012-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7776977
• 关键词: 3' Untranslated Regions; Acute; Addictive Behavior; Address; Age; Animals; Applied Research; Area; Atlases; Basic Science; Behavior; Behavioral; Binding; Binding Sites; Biological Assay; Biology; Brain; Brain region; Brain-Derived Neurotrophic Factor; Cell Line; Cells; Chimeric Proteins; Chronic; Cocaine; Cocaine Dependence; Consensus; Corpus striatum structure; DRD1 gene; Development; Dopamine; Dopamine D1 Receptor; Dopamine Receptor; Drug Addiction; Exhibits; Functional disorder; Gene Expression; Gene Expression Regulation; General Population; Genes; Goals; Hippocampus (Brain); In Vitro; Injection of therapeutic agent; Knockout Mice; Lead; Mediating; Messenger RNA; Methods; MicroRNAs; Molecular; Mus; National Institute of Neurological Disorders and Stroke; Nervous system structure; Neurons; Neurotransmitters; Nucleus Accumbens; Pathway interactions; Play; Post-Transcriptional Regulation; Process; Proteins; Receptor Gene; Regulation; Reporter; Reporter Genes; Reporting; Research; Rodent; Role; Self Administration; System; Testing; Transgenes; Transgenic Mice; Translational Research; Translations; Ventral Tegmental Area; cis acting element; experience; improved; mRNA Expression; novel; novel therapeutics; prevent; promoter; protein expression; public health relevance; receptor; receptor expression; research study; reward processing

DESCRIPTION (provided by applicant): The neurotransmitter dopamine plays an important role in the brain reward process. Dysfunction of the dopaminergic system is implicated in addictive behaviors. The relationship between dopaminergic system and cocaine addiction has been particularly well characterized. Numerous studies have shown that the D1 dopamine receptor subtype is involved in mediating the effects of cocaine. Acute and chronic administration of cocaine has been reported to alter the expression levels of D1 receptors in the mesocorticolimbic pathway. The molecular mechanisms and extra cellular factors that mediate the changes in dopamine receptor expression in cocaine-treated animals are largely unknown; however some reports have indicated that changes occur at the post transcriptional level. We have recently determined some of the extra cellular factors and molecular mechanisms that regulate the expression of the D1 dopamine receptor gene. Using a catecholaminergic neuronal cell line we determined that the endogenous D1 dopamine receptor is regulated at the post-transcriptional level during in vitro differentiation, and also following brain-derived neurotrophic factor treatment. Our preliminary studies also show that the post-transcriptional regulation of D1 dopamine receptor expression is mediated by cis-acting elements in the 3' untranslated region of the gene. In this R03 project, we test a novel hypothesis that posttranscriptional modulation of D1 dopamine receptor expression by acute and chronic administration of cocaine is mediated by microRNAs that bind cis-acting elements in the 3' untranslated region of the D1 receptor gene. To test this hypothesis, we will use the Drd1a-EGFP reporter transgenic mice generated by the NINDS Gene Expression Nervous System Atlas (GENSAT) project. Following acute and chronic cocaine treatment of Drd1a-EGFP reporter transgenic mice, we will evaluate the expression of D1 receptor mRNA and D1 receptor protein the nucleus accumbens, caudate, cortex, hippocampus and ventral tegmental area. In addition, we will also assay the expression of putative microRNAs that target the D1 receptor gene in the cocaine-treated animals from various brain regions. The goal of this R03 project is to determine if cocaine-induced changes in D1 dopamine receptor expression can be related to changes in expression of a specific microRNA, and to demonstrate, using primary cultures of D1 receptor-expressing neurons, that altering the expression of this microRNA will also alter the expression of D1 receptor protein. To date, there have been no reports of microRNAs regulating dopamine receptor expression. The results from this project would open a new area of research in dopamine receptor biology and improve our understanding of molecular mechanisms that regulate changes in dopamine receptor expression in cocaine addiction in particular, and addictive processes in general. PUBLIC HEALTH RELEVANCE: The goal of this proposal is to determine the role of microRNAs in mediating regulation of D1 dopamine receptor expression in cocaine addiction. The project will identify the microRNA that is involved and demonstrate how it mediates the post-transcriptional regulation of D1 receptor expression in cocaine-treated animals. The results of this proposal will open a new area of research in dopamine receptor biology and lead to the development of potentially novel therapeutic methods for treating cocaine addiction.

描述（由申请人提供）：神经递质多巴胺在大脑奖励过程中发挥着重要作用。多巴胺能系统的功能障碍与成瘾行为有关。多巴胺能系统与可卡因成瘾之间的关系已得到特别明确的描述。大量研究表明，D1 多巴胺受体亚型参与介导可卡因的作用。据报道，急性和慢性服用可卡因会改变皮质边缘通路中 D1 受体的表达水平。介导可卡因治疗动物多巴胺受体表达变化的分子机制和细胞外因素在很大程度上尚不清楚；然而，一些报告表明变化发生在转录后水平。我们最近确定了一些调节 D1 多巴胺受体基因表达的细胞外因子和分子机制。使用儿茶酚胺能神经元细胞系，我们确定内源性 D1 多巴胺受体在体外分化期间以及脑源性神经营养因子治疗后在转录后水平受到调节。我们的初步研究还表明，D1 多巴胺受体表达的转录后调节是由该基因 3' 非翻译区的顺式作用元件介导的。在这个 R03 项目中，我们测试了一个新的假设，即急性和慢性服用可卡因对 D1 多巴胺受体表达的转录后调节是由结合 D1 受体基因 3' 非翻译区顺式作用元件的 microRNA 介导的。为了验证这一假设，我们将使用 NINDS 基因表达神经系统图谱 (GENSAT) 项目生成的 Drd1a-EGFP 报告基因转基因小鼠。对 Drd1a-EGFP 报告基因转基因小鼠进行急性和慢性可卡因治疗后，我们将评估伏核、尾状核、皮质、海马和腹侧被盖区 D1 受体 mRNA 和 D1 受体蛋白的表达。此外，我们还将检测可卡因治疗动物不同脑区中靶向 D1 受体基因的假定 microRNA 的表达。该 R03 项目的目标是确定可卡因诱导的 D1 多巴胺受体表达变化是否与特定 microRNA 表达的变化有关，并利用表达 D1 受体的神经元的原代培养物证明，改变该 microRNA 的表达也会改变 D1 受体蛋白的表达。迄今为止，还没有关于 microRNA 调节多巴胺受体表达的报道。该项目的结果将开辟多巴胺受体生物学的新研究领域，并提高我们对调节可卡因成瘾（特别是可卡因成瘾）中多巴胺受体表达变化以及一般成瘾过程的分子机制的理解。 公共健康相关性：本提案的目标是确定 microRNA 在介导可卡因成瘾过程中 D1 多巴胺受体表达调节中的作用。该项目将鉴定所涉及的 microRNA，并展示它如何介导可卡因治疗动物中 D1 受体表达的转录后调节。该提案的结果将开辟多巴胺受体生物学的新研究领域，并导致开发治疗可卡因成瘾的潜在新颖治疗方法。