Modulation of frontal cortex NDRG1 and the effects on myelination, ethanol sensitivity, and ethanol consumption

额叶皮质 NDRG1 的调节及其对髓鞘形成、乙醇敏感性和乙醇消耗的影响

• 批准号: 9763391
• 负责人: Guy M Harris
• 金额: $ 3.37万
• 依托单位: VIRGINIA COMMONWEALTH UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-09-25 至 2020-08-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9763391
• 关键词: Acute; Adolescent; Affect; Alcohol consumption; Behavioral; Biological; Candidate Disease Gene; Cells; Charcot-Marie-Tooth Disease; Chronic; Consumption; Development; Disease; Dose; Down-Regulation; Ethanol; Event; Gene Expression; Gene Proteins; Genes; Genetic Determinism; Genomics; Glucocorticoids; Hippocampus (Brain); Human; Immunohistochemistry; Individual; Knockout Mice; Laboratories; Link; Manuscripts; Messenger RNA; Modeling; Molecular; Mus; Mutation; Myelin; Myelin Proteins; NDRG1 gene; Neoplasm Metastasis; Neurons; Oligodendroglia; Pathologic; Pathway interactions; Peripheral; Phenotype; Phosphorylation; Phosphotransferases; Physiological; Play; Predisposing Factor; Prefrontal Cortex; Preparation; Proteins; Reflex action; Regulation; Risk; Risk Factors; Rodent; Role; Schwann Cells; Site; Somatosensory Cortex; Stress; Structure; Time; Tissues; Transcript; Variant; Viral Vector; Work; alcohol behavior; alcohol effect; alcohol exposure; alcohol response; alcohol risk; alcohol sensitivity; alcohol use disorder; behavior measurement; behavioral response; cell type; drinking; drinking behavior; frontal lobe; hippocampal pyramidal neuron; individual response; knock-down; mRNA Expression; myelination; neuroimaging; overexpression; preference; problem drinker; protein expression; response; single-cell RNA sequencing; small hairpin RNA; transcriptome

Project Summary This project aims to study and better define a potential biological pathway which multiple pieces of evidence have suggested plays a role in behavioral responses to ethanol. This pathway is centered around N-myc downstream-regulated gene 1 (NDGR1), whose expression in the prefrontal cortex (PFC) has been linked the acute ethanol sensitivity phenotype, loss of righting reflex (LORR) duration. Mutations in NDRG1 in humans are the cause of Charcot-Marie-Tooth disease type 4D (CMT4D), a disease that severely affects peripheral myelination by inhibiting the function of Schwann Cells. In the CNS, NDRG1 expression has been shown to be a key network regulator of multiple myelin-related genes in the PFC, particularly in the presence of ethanol. Multiple studies have found that NDRG1 expression levels and sub-cellular localization varies substantially between cell-types throughout the body. Additionally, there is a large amount of evidence to suggest that phosphorylation of the protein alters its physiological function within the cell. Using a time course and dose response study, we will investigate the regulation of NDRG1 by ethanol and ethanol’s effects on the level of mRNA expression, protein abundance, protein phosphorylation, and protein localization within oligodendrocytes and pyramidal neurons of the PFC of mice after acute doses of ethanol and after a voluntary consumption drinking study. Additionally, we will develop, validate, and use viral vectors for cell-type specific overexpression NDRG1 in either oligodendrocytes or neurons within the PFC of mice. We will then study the effect of overexpression of this transcript on LORR duration and ethanol consumption and preference by providing voluntary ethanol access via an intermittent access drinking model. Finally, we will investigate the effect of overexpression of this transcript on basal myelin-related gene expression within the PFC and the overall transcriptome of the PFC.

项目摘要 该项目旨在研究和更好地定义一个潜在的生物途径， 有证据表明，在对乙醇的行为反应中起着重要作用。该途径 以N-myc下游调节基因1（NDGR 1）为中心，NDGR 1在 前额叶皮层（PFC）与急性乙醇敏感性表型、翻正能力丧失 反射（LORR）持续时间。人类NDRG 1突变是腓骨肌萎缩症的原因 疾病类型4D（CMT 4D），一种通过抑制外周髓鞘形成而严重影响外周髓鞘形成的疾病， 雪旺细胞的功能在中枢神经系统中，NDRG 1的表达已被证明是一个关键， PFC中多个髓鞘相关基因的网络调节因子，特别是在存在 乙醇多项研究发现，NDRG 1表达水平和亚细胞定位 在整个身体的细胞类型之间有很大的不同。此外，还有一个大的 大量的证据表明，蛋白质的磷酸化改变了其生理 在细胞内发挥作用。使用时间过程和剂量反应研究，我们将调查 乙醇对NDRG 1的调控及乙醇对NDRG 1 mRNA表达、蛋白表达水平的影响 丰度、蛋白磷酸化和蛋白定位， 在急性剂量的乙醇和自愿的 饮酒研究。此外，我们将开发，验证和使用病毒载体， 在PFC内的少突胶质细胞或神经元中的细胞类型特异性过表达NDRG 1 对小鼠然后我们将研究该转录本的过表达对LORR持续时间的影响， 乙醇消费和偏好，通过间歇性提供自愿乙醇获取， 饮酒模式。最后，我们将研究过表达该转录本的影响， 对PFC内基础髓鞘相关基因表达和PFC的整体转录组的影响。 PFC.