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

项目摘要 该项目旨在研究和更好地定义一条潜在的生物途径，该途径由多个片段组成 有证据表明，酒精在行为反应中起到了作用。这条路是 以N-myc下游调控基因1(NDGR1)为中心，其在 前额叶皮质(PFC)与急性酒精敏感性表型、翻正能力丧失有关 反射(LORR)持续时间。人类NDRG1基因突变是Charcot-Marie-Tooth的原因 4D型疾病(CMT4D)，一种通过抑制外周髓鞘形成而严重影响的疾病 雪旺细胞的功能。在CNS中，NDRG1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，特别是在存在 乙醇。多项研究发现，NDRG1的表达水平和亚细胞定位 在全身不同的细胞类型之间有很大的差异。此外，还有一个很大的 大量证据表明蛋白质的磷酸化改变了它的生理功能 在单元格内的功能。利用时间进程和剂量反应研究，我们将调查 乙醇对NDRG1基因表达的调节及乙醇对其基因表达和蛋白水平的影响 少突胶质细胞和少突胶质细胞内的丰度、蛋白磷酸化和蛋白定位 急性剂量乙醇和自愿给药后小鼠前额叶皮质锥体神经元的变化 饮酒消费研究。此外，我们将开发、验证和使用病毒载体来 前额叶内少突胶质细胞或神经元中NDRG1的细胞类型特异性过表达 老鼠的世界。然后我们将研究该转录本的过度表达对LOR持续时间和 通过间歇性提供自愿乙醇访问来实现乙醇消费和偏好 进入饮酒模式。最后，我们将调查该转录本过度表达的影响 前额叶髓鞘相关基因的基础表达及其转录组的研究 一等兵。

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.