Effects of Adenylyl Cyclases 1 and 8 on Neuronal Sensitivity to Ethanol

这项提案的长期目标是确定神经毒性效应的潜在机制。 酒精在胎儿酒精综合征(FAS)中的作用。腺酰环化酶AC1和/或ACS的缺失使其变得敏感 然而，在活动阻断后纹状体神经元发育至死亡，其神经保护机制 这些AC的行为是未知的。这项提议将检验AC1或ACS通过唯一方式起作用的假设 乙醇诱导NMDA受体信号转导的突触前和突触后机制 曝光或活动封锁。我们将通过以下目的来检验这一假设：i)定义解剖学 AC1和ACS与乙醇诱导的纹状体突触突触前、后的关系 利用免疫组织化学和生化分级技术诱导细胞凋亡；ii)测定 纹状体神经元对乙醇和NMDA受体拮抗剂活动阻断的敏感性及鉴定 AC1和ACS在常规和条件性AC缺失小鼠体内的生存蛋白靶点； 和iii)确定皮质纹状体共培养细胞对活动阻断的敏感性和分子水平 AC1和ACS调节这种敏感性的机制。定义分子机制和 决定纹状体和其他脑区对酒精神经毒性的敏感性或抵抗力的靶点 对于限制与Fas相关的病理性后遗症的治疗设计至关重要。 候选人目前是博士后研究员，他的职业目标是阐明 乙醇对神经元敏感性的作用，重点了解乙醇对神经细胞的毒性。 发育中的大脑。在华盛顿大学路易斯·穆格里亚博士的指导下进行的科学培训 这是一个理想的环境，让候选人能够获得成为独立调查员所需的技能。 作为她培训的主要部分，候选人将开发一种新的皮质纹状体共培养系统来解剖 AC1/AC8在体外的作用，在卡伦·奥马利博士的共同赞助下，他是这一领域的专家 多巴胺能信号和神经毒性的影响。使用共聚焦成像和活性染料的培训 将由史蒂夫·门纳里克博士提供。应聘者的目标是在此期间尽早获得教员职位。 在培训期间，并在奖励的最后几年申请独立资助。 与公共健康的相关性：也许饮酒最大的长期影响是对 发育中的胎儿，这可能导致胎儿酒精综合症。尽管与饮酒的联系在 妊娠和胎儿的神经毒性是明确的，其病理后果的潜在机制是 大脑仍然不确定，阻碍了改进疗法的发展。建议的目标是 研究项目是阐明与乙醇诱导相关的分子机制和靶点。 发育中的大脑中的神经毒性。

The long-term objectives of this proposal are to define the mechanisms underlying the neurotoxic effects of ethanol in Fetal Alcohol Syndrome (FAS). Deletion of adenylyl cyclases, AC1 and/or ACS,sensitizes developing striatal neurons to death after activity blockade, however, the neuroprotective mechanisms by which these ACs act are unknown. This proposal will test the hypothesis that AC1 or ACS act by unique presynaptic and postsynaptic mechanisms to increase NMDA receptor signaling in the setting of ethanol exposure or activity blockade. We will test this hypothesis with the following Aims: I) define the anatomic relationship of AC1 and ACS to pre- and postsynaptic aspects of striatal synapses undergoing ethanol- induced apoptosis using immunohistochemical and biochemical fractionation techniques; II) determine the sensitivity of striatal neurons to activity blockade by ethanol and NMDA receptor antagonists and identify prosurvival protein targets of AC1 and ACS in vivo using mice with conventional and conditional AC deletion; and III)determine the sensitivity to activity blockade of corticostriatal co-cultures and the molecular mechanisms by which AC1 and ACS modulate this sensitivity. Defining the molecular mechanisms and targets that determine sensitivity or resistance to ethanol neurotoxicity in the striatum and other brain regions is critical to the design of therapies to limit the pathological sequelea associated with FAS. The candidate is currently a postdoctoral fellow whose career goal is to elucidate the mechanisms of ethanol action on neuronal sensitivity, with an emphasis on understanding ethanol-induced neurotoxicity in the developing brain. Mentored scientific training under Dr. Louis Muglia at Washington University provides an ideal setting to do so, allowing the candidate to gain skills needed to become an independent investigator. As a major part of her training, the candidate will develop a novel corticostriatal co-culture system to dissect the roles of AC1/AC8 in vitro, under co-sponsorship of Dr. Karen O'Malley, an expert in the field of dopaminergic signaling and the effects of neurotoxicity. Training in the use of confocal imaging and vital dyes will be provided by Dr. Steve Mennerick. The candidate aims to achieve a faculty position early during this training period and apply for independent funding during the final years of the award. Relevance to Public Health: Perhaps the greatest long-term impact of alcohol use is the effects on the developing fetus, which can result in fetal alcohol syndrome. Though the association of alcohol use during pregnancy and fetal neurotoxicity is clear, mechanisms underlying the pathological consequences in the brain remain undefined, hindering the development of improved therapies. The goal of the proposed research project is to elucidate the molecular mechanisms and targets associated with ethanol-induced neurotoxicity in the developing brain.