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定义乙醇在胎儿酒精综合征（FAS）中的神经毒性作用的基础机制。腺苷酸环化酶AC1和/或AC的缺失使发展后的纹状体神经元在活动封锁后致死至死亡，但是，这些ACS作用的神经保护机制尚不清楚。该建议将检验以下假设：AC1或ACS通过独特的突触前和突触后机制作用，以增加乙醇暴露或活性阻滞的情况下增加NMDA受体信号传导。我们将以以下目的检验这一假设：i）定义AC1和ACS与使用免疫组织化学和生化分离化学的乙醇诱导凋亡的纹状体突触前和突触后方面的解剖学关系； ii）确定乙醇和NMDA受体拮抗剂对纹状体神经元对活性阻断的敏感性，并使用带有常规和条件AC缺失的小鼠在体内鉴定AC1和ACS的PROVIVAR蛋白靶标； iii）确定对皮质纹状体培养的活性阻滞的敏感性以及AC1和ACS调节这种敏感性的分子机制。定义确定纹状体和其他大脑区域中对乙醇神经毒性敏感性或抗性的分子机制和靶标对于限制与FAS相关的病理后遗症的疗法至关重要。候选人目前是博士后研究员，其职业目标是阐明乙醇对神经元敏感性的机制，重点是了解乙醇引起的神经毒性在发育中的大脑中。华盛顿大学路易斯·穆吉亚（Louis Muglia）的指导培训提供了理想的环境，使候选人能够获得成为独立调查员所需的技能。作为她培训的主要部分，候选人将开发一种新型的皮质乳化共培养系统，以剖析AC1/AC8在体外的作用，该系统是在多巴胺能信号传导领域的专家Karen O'Malley博士的共同赞助下，及其神经毒性的影响。史蒂夫·梅纳里克（Steve Mennerick）博士将提供有关共聚焦成像和重要染料的使用培训。候选人旨在在此培训期间早期获得教师职位，并在奖励的最后几年申请独立资金。