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酒精对胎儿酒精综合征(FAS)的神经毒性作用的潜在机制。腺酰环化酶AC1和/或ACS的缺失使发育中的纹状体神经元在活动阻断后对死亡敏感，然而，这些ACS作用的神经保护机制尚不清楚。这一提议将检验AC1或ACS通过独特的突触前和突触后机制在酒精暴露或活动阻断的背景下增加NMDA受体信号的假设。我们将通过下列目标来检验这一假说：i)使用免疫组织化学和生化分级技术确定AC1和ACS与乙醇诱导的纹状体突触的突触前和突触后方面的解剖关系；ii)确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，并使用常规和有条件AC缺失的小鼠在体内确定AC1和ACS的存活蛋白质靶点；iii)确定AC1和ACS对活动阻断的敏感性以及AC1和ACS调节这种敏感性的分子机制。确定决定纹状体和其他脑区对乙醇神经毒性的敏感性或抵抗力的分子机制和靶点，对于设计限制与Fas相关的病理后遗症的治疗方案至关重要。这位候选人目前是博士后，他的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·穆格里亚博士的指导下进行的科学培训为这样做提供了一个理想的环境，使候选人能够获得成为独立调查人员所需的技能。作为她的培训的主要部分，候选人将开发一种新的皮质纹状体共培养系统，在体外分析AC1/AC8的作用，由Karen O‘Malley博士共同赞助，Karen O’Malley博士是多巴胺能信号和神经毒性影响领域的专家。史蒂夫·门纳里克博士将提供使用共聚焦成像和活性染料的培训。候选人的目标是在培训期间尽早获得教员职位，并在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.