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

描述（由申请人提供）：本提案的长期目标是确定胎儿酒精综合征（FAS）中乙醇神经毒性作用的潜在机制。腺苷酸环化酶，AC 1和/或ACS的删除，敏感性发育中的纹状体神经元死亡后，活动封锁，然而，这些AC的神经保护机制是未知的。该提议将检验AC 1或ACS通过独特的突触前和突触后机制在乙醇暴露或活动阻断的情况下增加NMDA受体信号传导的假设。我们将用以下目的来检验这一假说：I）用免疫组织化学和生化分离技术确定AC 1和ACS与经历乙醇诱导的凋亡的纹状体突触的突触前和突触后方面的解剖学关系;（二）测定纹状体神经元对乙醇和NMDA受体拮抗剂的活性阻断的敏感性，并使用具有以下特征的小鼠在体内鉴定AC 1和ACS的促存活蛋白靶标：常规和条件性AC缺失;和III）确定皮质纹状体共培养物对活性阻断的敏感性以及AC 1和ACS调节该敏感性的分子机制。确定纹状体和其他脑区对乙醇神经毒性的敏感性或抗性的分子机制和靶点，对于设计治疗方案以限制FAS相关的病理后遗症至关重要。该候选人目前是一名博士后研究员，其职业目标是阐明乙醇对神经元敏感性的作用机制，重点是了解乙醇在发育中的大脑中诱导的神经毒性。在华盛顿大学路易斯·穆利亚博士的指导下进行的科学培训提供了一个理想的环境，使候选人能够获得成为独立调查员所需的技能。作为她的培训的一个主要部分，候选人将开发一种新的皮质纹状体共培养系统，在多巴胺能信号传导和神经毒性影响领域的专家Karen O 'Malley博士的共同赞助下，在体外剖析AC 1/AC 8的作用。共聚焦成像和活体染料的使用培训将由Steve Mennerick博士提供。候选人的目标是在培训期间尽早获得教职，并在该奖项的最后几年申请独立资助。

DESCRIPTION (provided by applicant): 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 sequelae 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.