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

腺苷酸环化酶 1 和 8 对神经元对乙醇敏感性的影响

• 批准号: 7512439
• 负责人: ALANA C. CONTI
• 金额: $ 10.75万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-09-20 至 2009-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7512439
• 关键词: Adenylate Cyclase; Alcohol-Induced Neurotoxicity; Anatomy; Apoptosis; Award; Biochemical; Brain; Brain region; Cell Death; Cessation of life; Chromosome Pairing; Coculture Techniques; Corpus striatum structure; Dose; Dyes; Ethanol; Faculty; Fetal Alcohol Syndrome; Fractionation; Funding; Genetic; Goals; Image; In Vitro; Ketamine; Localized; Measures; Mentors; Molecular; Mus; N-Methyl-D-Aspartate Receptors; NMDA receptor antagonist; Neurons; Positioning Attribute; Postdoctoral Fellow; Proteins; Receptor Signaling; Research Personnel; Resistance; Role; Signal Pathway; Signal Transduction; Signaling Protein; Synapses; Synaptic Vesicles; System; Techniques; Testing; Training; Universities; Washington; adenylyl cyclase 1; alcohol effect; alcohol exposure; alcohol sensitivity; career; in vivo; intracellular protein transport; neurotoxic; neurotoxicity; novel; postsynaptic; presynaptic; protein localization location; response; skills; therapy design

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.

描述（由申请人提供）：该提案的长期目标是确定乙醇对胎儿酒精综合症（FAS）的神经毒性作用的机制。腺苷酸环化酶、AC1 和/或 ACS 的缺失会使发育中的纹状体神经元在活动阻断后变得敏感而死亡，然而，这些 AC 发挥作用的神经保护机制尚不清楚。该提案将检验以下假设：AC1 或 ACS 通过独特的突触前和突触后机制发挥作用，在乙醇暴露或活动阻断的情况下增加 NMDA 受体信号传导。我们将通过以下目标来检验这一假设：I) 使用免疫组织化学和生化分级分离技术，定义 AC1 和 ACS 与经历乙醇诱导细胞凋亡的纹状体突触前和突触后方面的解剖关系； II) 使用常规和条件 AC 缺失的小鼠，确定纹状体神经元对乙醇和 NMDA 受体拮抗剂活性阻断的敏感性，并在体内鉴定 AC1 和 ACS 的促存活蛋白靶点； III) 确定皮质纹状体共培养物对活性阻断的敏感性以及 AC1 和 ACS 调节这种敏感性的分子机制。确定纹状体和其他大脑区域对乙醇神经毒性的敏感性或抵抗力的分子机制和靶标对于设计限制与 FAS 相关的病理后遗症的疗法至关重要。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引起的神经毒性。华盛顿大学 Louis Muglia 博士指导的科学培训为此提供了理想的环境，使候选人能够获得成为独立调查员所需的技能。作为培训的主要部分，候选人将在多巴胺能信号传导和神经毒性影响领域专家 Karen O'Malley 博士的共同资助下，开发一种新型皮质纹状体共培养系统，以在体外剖析 AC1/AC8 的作用。 Steve Mennerick 博士将提供共焦成像和活体染料使用方面的培训。候选人的目标是在培训期间尽早获得教职，并在奖项的最后几年申请独立资助。