Regulation of alcohol consumption by chromatin modification

通过染色质修饰调节饮酒

• 批准号: 8527622
• 负责人: Paula Hoffman
• 金额: $ 32.52万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-09-30 至 2014-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8527622
• 关键词: Acetylation; Affect; Alcohol consumption; Alcohol dependence; Alcoholism; Alcohols; Amygdaloid structure; Autacoids; Binding; Biological Assay; Bispecific Antibody 2B1; Brain; Candidate Disease Gene; Cell Communication; Cell Nucleus; Characteristics; Chromatin; Chronic; Cocaine; Code; Cytoplasm; Cytosol; DBA/2 Mouse; Data; Dependence; Development; Enzymes; Ethanol; GTP-Binding Proteins; Gene Expression; Gene Proteins; Genes; Genetic; Genetic Transcription; Genomics; HDAC4 gene; HDAC5 gene; Heavy Drinking; Histone Acetylation; Histones; Immunoblotting; Immunohistochemistry; Inbred Strains Mice; Inbreeding; Injection of therapeutic agent; Lead; Lentivirus Vector; Link; Literature; Mass Spectrum Analysis; Measures; Mediating; Microarray Analysis; Modification; Molecular; Monitor; Mouse Strains; Mus; Neurosciences; Nuclear; Nucleus Accumbens; Pharmaceutical Preparations; Phenotype; Predisposition; Promoter Regions; Proteins; RNA Interference; Rattus; Reaction; Recombinant Inbred Strain; Regulation; Relative (related person); Reporting; Rewards; Rodent; Role; Signal Pathway; Techniques; Transcript; Variant; Viral Vector; base; chromatin modification; dimer; drinking; histone modification; human GNB1 protein; inhibitor/antagonist; novel strategies; phenomics; preference; promoter; response; small hairpin RNA

DESCRIPTION (provided by applicant): During the previous INIA (Integrated Neurosciences Initiative on Alcoholism)-West project period a genetical genomic/phenomic approach was applied to identify candidate genes in rodents that, through variation in expression levels, contribute to the predisposition to consume varying amounts of alcohol. One such candidate gene is Gnb1 (the G protein beta 1 subunit, Gpi). Brain levels of the Gpi protein are inversely correlated with levels of alcohol consumption in inbred and recombinant inbred strains of mice and in lines of mice selected for differences in alcohol consumption. Treatment of mice with a lentiviral vector expressing a shRNA targeting Gnb1 lowered Gpi levels in nucleus accumbens and increased alcohol consumption by DBA/2 mice. Gpi, as a dimer with Gy proteins, affects numerous intracellular signaling pathways, and has also been reported to interact with, and modify the transcriptional activity of, the Class lla histone deacetylases (HDACs) 4 and 5. HDACs generally repress transcription via modification of histone proteins and class lla HDACs can shuttle between nucleus and cytoplasm, which can regulate their transcriptional activity. In particular, the direct Gpy-HDAC interaction reduces HDAC activity, and therefore has the potential to regulate the transcription of other genes. Given that HDACS activity has previously been found to be crucial for the rewarding effect of cocaine, and that HDAC inhibition reduces alcohol consumption, a role for the Gpy-HDAC interaction in alcohol consumption is postulated. Alcohol consumption will be measured after lowering Gnb1 expression in brains of low alcohol-consuming mice, and after inhibiting or lowering expression of HDACs in brain of high alcohol-consuming mice. To assess the link between Gpi and HDACs, HDAC transcriptional activity (transcript levels and promoter acetylation), as well as HDAC subcellular localization, will be determined after lowering Gpi with RNAi. Overall, this project will investigate molecular mechanisms, focusing on chromatin modification, by which a previously identified candidate gene for alcohol consumption/preference influences transcriptional networks that may underlie this phenotype.

描述（由申请人提供）：在之前的 INIA（酒精中毒综合神经科学倡议）-West 项目期间，应用遗传基因组/表型组学方法来识别啮齿类动物中的候选基因，这些基因通过表达水平的变化导致不同量酒精消耗的倾向。其中一个候选基因是 Gnb1（G 蛋白 β 1 亚基，Gpi）。在近交系和重组近交系小鼠以及因酒精消耗差异而选择的小鼠品系中，Gpi蛋白的脑水平与酒精消耗水平呈负相关。用表达针对 Gnb1 的 shRNA 的慢病毒载体治疗小鼠，可降低伏隔核中的 Gpi 水平，并增加 DBA/2 小鼠的饮酒量。 Gpi，作为与Gy蛋白的二聚体，影响许多细胞内信号传导途径，并且还据报道与IIa类组蛋白脱乙酰酶(HDAC)4和5相互作用并改变其转录活性。HDAC通常通过组蛋白的修饰来抑制转录，并且IIa类HDAC可以在细胞核和细胞质之间穿梭，这可以调节它们的转录 活动。特别是，Gpy-HDAC 的直接相互作用会降低 HDAC 活性，因此有可能调节其他基因的转录。鉴于先前已发现 HDACS 活性对于可卡因的奖赏效果至关重要，并且 HDAC 抑制可减少酒精消耗，因此推测 Gpy-HDAC 相互作用在酒精消耗中的作用。降低低度饮酒小鼠大脑中的 Gnb1 表达，以及抑制或降低高度饮酒小鼠大脑中 HDAC 的表达后，测量酒精消耗量。为了评估 Gpi 和 HDAC 之间的联系，将在用 RNAi 降低 Gpi 后确定 HDAC 转录活性（转录本水平和启动子乙酰化）以及 HDAC 亚细胞定位。总体而言，该项目将研究分子机制，重点关注染色质修饰，通过这种修饰，先前确定的饮酒/偏好候选基因会影响可能构成该表型的转录网络。