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.
It has long been thought that the initial sensitivity of an individual?s response to acute ethanol plays a significant role in determining the long-term risk that individual has of developing an Alcohol Use Disorder (AUD). Recent evidence in our lab suggests that N-myc downstream- regulated gene 1 (NDRG1) expression in the prefrontal cortex (PFC) is a key determinant of rodent sensitivity to acute ethanol phenotypes. We aim to characterize the regulation of ethanol exposure on NDRG1 expression, phosphorylation, and localization within the cells of the PFC and to determine if modulation of NDRG1 expression can affect acute ethanol phenotypes, thus potentially altering the long-term risk of AUD development, which will be investigated by studying if modulation alters ethanol drinking behavior.
