7/8 NADIA U01 Recovery of Adolescent Alcohol Disruption of Basal Forebrain-Cortical Projection Circuits

C7/8 UO 1：丽莎萨维奇和瑞安维特雷诺 青少年酒精性基底前脑-皮质投射回路破坏的恢复 项目摘要 青春期大量饮酒与大脑结构的持续变化有关， 连接和成人皮质介导的认知功能。持续观察到严重病理学变化 青少年间歇性乙醇暴露（Adolescent Intermittent Ethanol; AIE） 皮质神经生长因子（NGF），抑制胆碱能表型，以及长期减少 功能性胆碱能基底前脑神经元（CBFN）的数量。胆碱能投射神经元 在基底核大细胞复合体（NbM）内向额叶皮质提供乙酰胆碱（ACh）， 包括内侧前额叶皮质（mPFC）和眶额皮质（OFC）。行为刺激外排 AIE后，mPFC和OFC中的ACh均变钝， 灵活性.我们最近的数据表明，AIE诱导的CBFN丢失是由于表观遗传沉默， 胆碱能表型标记，但运动，可能通过神经生长因子机制，可以逆转这些 表观遗传变化和拯救胆碱能神经元。这表明，AIE最初是由一个 减少胆碱能神经元表型的表达，而不是神经元细胞死亡。因此，在本发明中， 应探讨AIE后CFBN病理恢复的机制。我们的首要假设是 AIE诱导的皮质投射部位NGF减少导致胆碱能神经元的表观遗传沉默 表型与NbM，回缩和/或功能障碍的神经再支配的皮质-NbM胆碱能 连接体，导致皮质胆碱能张力减弱和认知功能障碍。然而，适当地 AIE后的定时基于NGF的治疗（运动，NGF基因治疗，CRISPR/dCas 9-P300编辑）将 通过胆碱能基因的振兴来重振胆碱能前脑回路，这将挽救 皮质ACh，并恢复AIE诱导的皮质依赖性行为损伤。我们的目标是绘制地图， 挽救AIE后出现的胆碱能前脑和皮质回路病理（目的1），恢复认知能力 通过运动、神经生长因子基因治疗或选择性化学刺激胆碱能神经元发挥功能（目的 2），并通过CRISPR/dCas 9编辑，确定NGF缺陷在AIE诱导的表观遗传中的核心作用。 胆碱能表型基因的沉默（Aim 3）。越来越多的证据支持神经保护作用， NGF作为一个过程，以拯救脆弱的CBFN经历神经变性，我们有强有力的证据， AIE诱导的胆碱能病理是可以逆转的。了解胆碱能的机制 神经元的恢复将有助于开发更有效的治疗认知功能障碍的疗法 与酒精相关的脑损伤和其他神经退行性疾病有关。

C7/8 UO1: Lisa Savage & Ryan Vetreno Recovery of Adolescent Alcohol Disruption of Basal Forebrain-Cortical Projection Circuits Project Summary Heavy alcohol consumption during adolescence is associated with persistent changes in brain structure, connectivity, and adult cortical-mediated cognitive function. Critical pathological changes consistently observed in rodent models of adolescent binge ethanol exposure (Adolescent Intermittent Ethanol; AIE) are a reduction of cortical nerve growth factor (NGF), suppression of the cholinergic phenotypes, and a long-term decrease in the number of functional cholinergic basal forebrain neurons (CBFNs). The cholinergic projection neurons within the nucleus basalis magnocellularis complex (NbM) provide acetylcholine (ACh) to the frontal cortex, including the medial prefrontal cortex (mPFC) and orbitofrontal cortex (OFC). Behaviorally-stimulated efflux of ACh in both the mPFC and OFC are blunted following AIE, and there is a loss of cognitive and behavioral flexibility. Our recent data demonstrate that AIE-induced loss of CBFNs is due to epigenetic silencing of cholinergic phenotypic markers, but exercise, potentially through a NGF mechanism, can reverse these epigenetic changes and rescue cholinergic neurons. This demonstrates that AIE is initially marked by a reduction of the expression of cholinergic neuronal phenotypes, rather than neuronal cell death. Thus, mechanisms should be explored to recover CFBN pathology following AIE. Our overarching hypothesis is that AIE-induced reductions of NGF in cortical projection sites leads to epigenetic silencing of cholinergic phenotypes with the NbM, retraction and/or dysfunctional re-innervation of the cortical-NbM cholinergic connectome, causing blunting of cortical cholinergic tone and cognitive dysfunction. However, appropriately timed NGF-based therapies (exercise, NGF gene therapy, CRISPR/dCas9-P300 editing) following AIE will reinvigorate cholinergic forebrain circuitry through the revitalization of cholinergic genes, which will rescue cortical ACh and recover AIE-induced impairments in cortical-dependent behaviors. Our goals are to map and rescue cholinergic forebrain and cortical circuit pathology seen following AIE (Aim 1), restore cognitive functioning by exercise, NGF-gene therapy, or selective chemogenetic stimulation of cholinergic neurons (Aim 2), and identify the central role of NGF deficits, through CRISPR/dCas9 editing, in AIE-induced epigenetic silencing of cholinergic phenotype genes (Aim 3). Mounting evidence supports the neuroprotective effects of NGF as a course to rescue vulnerable CBFNs undergoing neurodegeneration, and we have strong evidence that AIE-induced cholinergic pathology can be reversed. Understanding the mechanisms of cholinergic neuronal recovery will aid in the development of more effective therapies to treat cognitive dysfunction associated with alcohol-related brain damage and other neurodegenerative disorders.