Interactive effects of ethanol and estrogen on brain vasopressin during puberty

描述（由申请人提供）：滥用酒精的女性患焦虑症的可能性是男性的两倍，这种现象的潜在生物学机制尚不清楚。我们的总体目标是确定酒精和雌激素对精氨酸抗利尿激素（AVP）的相互作用，AVP是一种公认的焦虑的关键分子介质，以阐明导致女性焦虑障碍风险增加的分子机制。青少年酗酒是成人焦虑症的潜在危险因素，因为青春期雌激素循环增加直接导致应激反应增强。青春期是大脑发育的一个动态而重要的时期，涉及皮质灰质、突触连通性的变化和神经发生的增加，对于青春期饮酒的长期神经生物学后果知之甚少。在这个广泛的大脑重塑的关键时期接触酒精可能导致永久性的神经元损伤或新神经元连接形成的中断，这可能表现为成人行为精神病，包括焦虑症。我们的初步数据显示：1)青春期饮酒会增加大脑特定区域AVP基因的表达。因此，在Specific Aim 1中提出的实验将直接验证以下假设：在青春期发育期间，AVP系统最容易受到酒精的影响；(2)雌激素加剧了酒精对AVP基因表达的影响。此外，我们的初步数据表明，酒精治疗和雌激素受体配体增加了源自下丘脑的神经细胞中AVP基因的表达，并且基因表达与基因启动子的活性密切相关。酒精还激活大脑中的雌激素信号通路，这表明酒精诱导AVP变化的潜在机制可能是由雌激素信号通路介导的。因此，在Specific Aim 2中提出的实验将直接验证以下假设：(1)急性酒精暴露会增加神经元细胞中AVP启动子的活性，(2)AVP启动子的特定调控区域与酒精相互作用，以及(3)雌激素和酒精协同作用增加AVP启动子活性。迄今为止，尚未确定青春期酒精激活的特定分子和神经内分泌标志物。这一建议的重点是一个特定的候选基因（AVP）及其下游信号通路在青春期发育形成。我们期望这些研究表明AVP是永久性改变的，要么作为酒精的直接靶点，要么通过类固醇激素受体信号通路间接改变。因此，这项研究的价值在于针对特定基因的治疗方法的潜力，也许可以逆转青春期发育期间饮酒引起的脑损伤，并加强在此期间戒酒的理由。滥用酒精的妇女患焦虑症的可能性是男性的两倍，这一现象的潜在生物学机制尚不清楚。这一建议的重点是一个特定的候选基因（AVP）及其下游信号通路在青春期发育形成。我们期望这些研究表明AVP是永久性改变的，要么作为酒精的直接靶点，要么通过类固醇激素受体信号通路间接改变。因此，这项研究的价值在于针对特定基因的治疗方法的潜力，也许可以逆转青春期发育期间饮酒引起的脑损伤，并加强在此期间戒酒的理由。

DESCRIPTION (provided by applicant): Women who abuse alcohol are twice as likely to develop anxiety disorders compared with men, a phenomenon in which the underlying biological mechanisms are unknown. Our overall objective is to identify the interactive effects of alcohol and estrogen on arginine vasopressin (AVP), a well-established key molecular mediator of anxiety, in order to elucidate the molecular mechanisms predisposing women to increased risk of anxiety disorders. Adolescent binge drinking is a potential risk factor for the development of adult anxiety disorders due to the heightened stress reactivity that occurs as a direct result of increased circulating estrogens during pubertal development. Little is known about the long-term neurobiological consequences of alcohol consumption during puberty, which is a dynamic and important period of brain development that involves changes in cortical gray matter, synaptic connectivity, and increased neurogenesis. Exposure of alcohol during this critical period of extensive brain remodeling may result in permanent neuronal damage or disruptions in the formation of new neuronal connections, which might manifest as adult behavioral psychoses, including anxiety disorder. Our preliminary data show that 1) alcohol exposure during puberty increased AVP gene expression in specific regions of the brain. Therefore, the experiments proposed in Specific Aim 1 will directly test the hypotheses that there is a critical window of time during pubertal development when the AVP system is most vulnerable to the effects of alcohol and (2) that estrogen exacerbates the effects of alcohol on AVP gene expression. Also, our preliminary data demonstrate that alcohol treatment and estrogen receptor ligands increased AVP gene expression in neuronal cells derived from the hypothalamus, and gene expression is closely correlated with the activity of the gene promoter. Alcohol also activates estrogen-signaling pathways in the brain, which suggests that the underlying mechanisms for alcohol-induced changes in AVP may be mediated by estrogen signaling pathways. Therefore, the experiments proposed in Specific Aim 2 will directly test the hypotheses that (1) acute alcohol exposure increases AVP promoter activity in neuronal cells, (2) that there are specific regulatory regions of the AVP promoter that interact with alcohol, and (3) that estrogen and alcohol interact synergistically to increase AVP promoter activity. To date, specific molecular and neuroendocrine markers that are activated by alcohol during puberty have not been identified. This proposal is focused on a specific candidate gene (AVP) and its downstream signaling pathways that are developmentally shaped during puberty. We expect these studies to show that AVP is permanently altered, either as a direct target for alcohol or indirectly through steroid hormone receptor signaling pathways. Thus, the value of this research lies in the potential for therapeutic approaches that would target specific genes and perhaps reverse brain damage caused by alcohol consumption during pubertal development as well as strengthen reasons for abstaining from alcohol during that time. PUBLIC HEALTH RELEVANCE Women who abuse alcohol are twice as likely to develop anxiety disorders compared with men, a phenomenon in which the underlying biological mechanisms are unknown. This proposal is focused on a specific candidate gene (AVP) and its downstream signaling pathways that are developmentally shaped during puberty. We expect these studies to show that AVP is permanently altered, either as a direct target for alcohol or indirectly through steroid hormone receptor signaling pathways. Thus, the value of this research lies in the potential for therapeutic approaches that would target specific genes and perhaps reverse brain damage caused by alcohol consumption during pubertal development as well as strengthen reasons for abstaining from alcohol during that time.