Medial Prefrontal Cortical Gliogenesis and Alcohol Dependence

项目概要/摘要 复发定义为长期戒酒后恢复饮酒， 酗酒是一个普遍而重要的公共卫生问题。复发是一个主要障碍， 治疗工作。一种有希望的方法，用于降低中度抑郁症受试者的高复发倾向 严重酒精使用障碍（AUD）是识别大脑中的脆弱性因素， 增强酒精饮用复发;内侧前额叶皮层（mPFC）是关键的大脑区域之一 与酒精饮酒行为的复发有关 我们使用慢性间歇性乙醇蒸汽吸入（CIE）模型的乙醇依赖， 表明雌性大鼠在CIE期间增加乙醇自我给药预示着更高的复发率， 与男性相比，在强迫禁欲过程中由乙醇上下文线索引发的乙醇饮用。在 此外，我们已经表明，复发期间饮酒与内皮细胞特异性 在两种性别的mPFC中均存在血管生成肽PECAM-1，表明血脑屏障（BBB）不稳定。 用抗血管生成剂治疗使两种性别的PECAM-1表达正常化并减少复发 酒精摄入量的变化 为了进一步了解BBB不稳定和功能障碍的机制，我们已经开始研究 通过内皮细胞富集试验从mPFC中分离内皮细胞。我们已经进行了RNA 测序以初步分析从未经乙醇处理的成年女性分离的细胞的内皮转录组， 雄性大鼠我们的研究结果表明，女性的内皮细胞有更高的基因表达， 血管生成和内皮细胞的稳定性，而男性细胞的基因表达更高， 与免疫反应有关。我们已经发表了从细胞因子和趋化因子多重 分析表明，在未经乙醇处理的成年雄性大鼠中，mPFC具有较高的前-和抗- 炎症细胞因子和趋化因子。这些研究结果支持了 对照、未处理乙醇条件下mPFC中的内皮转录组和免疫应答 以及在乙醇依赖期间它们是如何改变的开放性问题。 为了进一步支持血脑屏障的不稳定性，我们发现粘附连接蛋白钙粘蛋白5（Cdh 5，VE-1）在血脑屏障中表达。 钙粘蛋白）在酒精依赖性雌性和雄性大鼠的mPFC中显著改变。另外还 表明Cdh 5在急性戒断期间增加，在长期禁欲期间显著降低， 在两种性别中，与乙醇初治对照组相比，酒精饮用复发后增加。在 除了mPFC中的内皮变化外，电生理学研究表明， 酒精依赖性雌性和雄性大鼠的可塑性。因此，在更新申请中，我们希望定义 Cdh 5在戒酒期间通过改变酒精摄入量促进男女酒精摄入复发中的作用。 内皮转录组、促炎和抗炎反应以及mPFC中的神经元可塑性。我们 假设将通过3个具体目标进行检验。

Project summary/Abstract Relapse defined as the resumption of alcohol drinking following a prolonged period of abstinence, represents a prevalent and significant public health concern in alcoholism. Relapse is a major impediment to treatment efforts. One promising approach for reducing high propensity for relapse in subjects with moderate to severe alcohol use disorder (AUD) is the identification of vulnerability factors in the brain that contribute to enhanced relapse to ethanol drinking; the medial prefrontal cortex (mPFC) is one of the key brain regions implicated in relapse to ethanol drinking behaviors. We have used the chronic intermittent ethanol vapor inhalation (CIE) model of ethanol dependence to demonstrate that increased ethanol self-administration during CIE in female rats predicts higher relapse to ethanol drinking triggered by ethanol contextual cues during forced abstinence compared with males. In addition, we have shown that drinking during relapse is associated with increases in endothelial cell specific angiogenesis peptide PECAM-1 in the mPFC in both sexes, indicating blood-brain barrier (BBB) instability. Treatment with an anti-angiogenic agent normalized PECAM-1 expression in both sexes and reduced relapse to ethanol drinking in female rats. To further understand the mechanisms underlying BBB instability and dysfunction, we have begun studies to isolate endothelial cells from the mPFC via endothelial cell enrichment assay. We have performed RNA sequencing to initially profile endothelial transcriptome of cells isolated from ethanol naïve adult female and male rats. Our results show that endothelial cells in females have higher expression of genes associated with angiogenesis and endothelial cell stability, whereas cells from males have higher expression of genes associated with immune responses. We have published findings from cytokine and chemokine multiplex analyses to show that the mPFC in ethanol naïve adult male rats has higher expression of pro- and anti- inflammatory cytokines and chemokines compared with females. These findings support sexual dimorphism in the endothelial transcriptome and immune responses in the mPFC under control, ethanol naïve conditions and open questions on how they are altered during ethanol dependence. In further support of BBB instability, we show that adherens junction protein cadherin5 (Cdh5, VE- cadherin) is significantly altered in the mPFC in ethanol dependent female and male rats. Specifically we show that Cdh5 is increased during acute withdrawal, significantly decreased during prolonged abstinence, and increased following relapse to ethanol drinking compared to ethanol naïve controls in both sexes. In addition to the endothelial changes in the mPFC, electrophysiological studies demonstrate altered synaptic plasticity in ethanol dependent female and male rats. Therefore, in the renewal application we hope to define the role of Cdh5 during abstinence in promoting relapse to ethanol drinking in both sexes via altering the endothelial transcriptome, pro- and anti-inflammatory responses and neuronal plasticity in the mPFC. Our hypotheses will be tested via 3 Specific Aims.