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: 8321076
• 负责人: ALANA C. CONTI
• 金额: $ 11.19万
• 依托单位: WAYNE STATE 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 2014-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8321076
• 关键词: Adenylate Cyclase; Alcohol consumption; Alcohol-Induced Neurotoxicity; Anatomy; Apoptosis; Award; Biochemical; Brain; Brain region; Cell Death; Cessation of life; Coculture Techniques; Corpus striatum structure; Development; Dose; Dyes; Ethanol; Faculty; Fetal Alcohol Syndrome; Fetus; Fractionation; Funding; Genetic; Goals; Image; In Vitro; Ketamine; Measures; Mentors; Molecular; Mus; N-Methyl-D-Aspartate Receptors; NMDA receptor antagonist; Neurons; Positioning Attribute; Postdoctoral Fellow; Pregnancy; Proteins; Public Health; Receptor Signaling; Research Personnel; Research Project Grants; 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; fetal; improved; in vivo; neurotoxic; neurotoxicity; novel; postsynaptic; presynaptic; response; skills; therapy design

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.

这项建议的长期目标是确定神经毒性作用的机制 胎儿酒精综合征（FAS）腺苷酸环化酶AC 1和/或ACS的缺失使 发育中的纹状体神经元在活动阻断后死亡，然而， 这些进化人的具体行为尚不清楚该提议将检验AC 1或ACS通过独特的 乙醇环境中增加NMDA受体信号传导的突触前和突触后机制 暴露或活动封锁。我们将测试这一假设与以下目的：I）定义解剖 AC 1和ACS与纹状体突触的突触前和突触后方面的关系， 使用免疫组织化学和生物化学分级技术诱导细胞凋亡; II）确定 纹状体神经元对乙醇和NMDA受体拮抗剂活性阻断敏感性及鉴定 使用具有常规和条件性AC缺失的小鼠体内AC 1和ACS的促存活蛋白靶标; 和III）测定皮质纹状体共培养物对活性阻断的敏感性和分子生物学活性， AC 1和ACS调节这种敏感性的机制。定义分子机制， 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靶点 对于设计治疗以限制与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，重点是了解乙醇诱导的神经毒性， 大脑发育。在华盛顿大学路易斯·穆利亚博士的指导下， 这是一个理想的环境，让候选人获得成为独立调查员所需的技能。 作为她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， AC 1/AC 8在体外的作用，在Karen O 'Malley博士的共同赞助下， 多巴胺能信号传导和神经毒性的影响。共聚焦成像和活体染料使用培训 将由史蒂夫·门纳里克博士提供候选人的目标是在此期间尽早获得教职 培训期间，并在该奖项的最后几年申请独立的资金。 与公共卫生的相关性：也许酒精使用的最大长期影响是对健康的影响。 发育中的胎儿，这可能导致胎儿酒精综合征。尽管饮酒与 妊娠和胎儿神经毒性是明确的，机制的病理后果， 大脑仍然不确定，阻碍了改进疗法的发展。建议的目标 研究项目是阐明与乙醇诱导的 对大脑发育的神经毒性

项目成果

Dysregulation of TrkB phosphorylation and proBDNF protein in adenylyl cyclase 1 and 8 knockout mice in a model of fetal alcohol spectrum disorder.

胎儿酒精谱系障碍模型中腺苷酸环化酶 1 和 8 敲除小鼠中 TrkB 磷酸化和 proBDNF 蛋白的失调。

• DOI: 10.1016/j.alcohol.2015.11.008
• 发表时间: 2016
• 期刊: Alcohol (Fayetteville, N.Y.)
• 作者: Susick,LauraL;Chrumka,AlexandriaC;Hool,StevenM;Conti,AlanaC
• 通讯作者: Conti,AlanaC

Postnatal ethanol exposure simplifies the dendritic morphology of medium spiny neurons independently of adenylyl cyclase 1 and 8 activity in mice.

出生后乙醇暴露简化了小鼠中型多棘神经元的树突形态，与腺苷酸环化酶 1 和 8 活性无关。

• DOI: 10.1111/acer.12383
• 发表时间: 2014
• 期刊: Alcoholism, clinical and experimental research
• 作者: Susick,LauraL;Lowing,JenniferL;Provenzano,AnthonyM;Hildebrandt,ClaraC;Conti,AlanaC
• 通讯作者: Conti,AlanaC

Adenylyl cylases 1 and 8 mediate select striatal-dependent behaviors and sensitivity to ethanol stimulation in the adolescent period following acute neonatal ethanol exposure.

• DOI: 10.1016/j.bbr.2014.04.031
• 发表时间: 2014-08-01
• 期刊: BEHAVIOURAL BRAIN RESEARCH
• 影响因子: 2.7
• 作者: Susick, Laura L.;Lowing, Jennifer L.;Bosse, Kelly E.;Hildebrandt, Clara C.;Chrumka, Alexandria C.;Conti, Alana C.
• 通讯作者: Conti, Alana C.