Medial Prefrontal Cortical Gliogenesis and Alcohol Dependence

项目摘要/摘要 复发的定义是在长期戒酒后恢复饮酒， 在酒精中毒中代表着一个普遍和重大的公共卫生问题。旧病复发是一大障碍。 治疗方面的努力。一种有希望的方法来降低中度癌症患者的高复发倾向 重度酒精使用障碍(AUD)是对大脑中导致酒精使用障碍的易感因素的识别 内侧前额叶皮质(MPFC)是大脑的关键区域之一 与复吸酒精饮酒行为有关。 我们使用酒精依赖的慢性间歇性乙醇蒸气吸入(CIE)模型 在雌性大鼠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，在强制戒酒过程中，由酒精情境线索引发的酒精饮酒。在……里面 此外，我们还表明，复发期间饮酒与内皮细胞特异性增加有关。 血管生成多肽PECAM-1在mPFC中表达，提示血脑屏障(BBB)不稳定。 抗血管生成剂治疗使PECAM-1在两性中的表达正常化并减少复发 对雌性大鼠饮用酒精。 为了进一步了解血脑屏障不稳定和功能障碍的机制，我们已经开始研究 通过内皮细胞富集法从mPFC中分离内皮细胞。我们已经进行了核糖核酸 乙醇幼稚成年雌性和雌性乙醇分离细胞的内皮转录组初步测序 雄鼠。我们的结果表明，女性内皮细胞有较高的相关基因表达 血管生成和内皮细胞稳定性，而男性细胞有更高的基因表达 与免疫反应有关。我们已经发表了细胞因子和趋化因子复合体的研究结果。 分析表明，酒精未成熟的成年雄性大鼠的mPFC具有较高的正反向表达 炎性细胞因子和趋化因子与女性比较。这些发现支持人类的性二型性 对照、无酒精条件下mPFC内皮细胞转录组和免疫反应 以及关于它们在酒精依赖期间是如何改变的悬而未决的问题。 为了进一步支持血脑屏障的不稳定性，我们发现粘着连接蛋白钙粘蛋白5(CDH5，VE- 在酒精依赖的雌性和雄性大鼠的mPFC中，钙粘附素)显著改变。具体来说，我们 显示在急性戒断期间，CDH5升高，在长期戒断时显著降低， 与酒精幼稚的对照组相比，在再次饮酒后，两性的酒精摄入量都有所增加。在……里面 除了mPFC内皮细胞的变化外，电生理学研究还表明突触发生了变化 乙醇依赖雌性和雄性大鼠的可塑性。因此，我们希望在续签申请中定义 戒酒过程中CDH5在促进饮酒复吸中的作用 MPFC中的内皮转录组、促炎和抗炎反应以及神经元的可塑性。我们的 假说将通过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.