AC1 and AC8 as Mediators of the Dendritic Response to Ethanol in a Model of FASD

AC1 和 AC8 作为 FASD 模型中乙醇树突反应的介体

• 批准号: 8254943
• 负责人: Laura Susick
• 金额: $ 4.84万
• 依托单位: WAYNE STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-19 至 2014-09-18
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8254943
• 关键词: Acute; Adenylate Cyclase; Adult; Affect; Alcohol consumption; Alcohol-Induced Neurotoxicity; Area; Behavioral; Biological Process; Brain; Brain region; Brain-Derived Neurotrophic Factor; Calcium; Cell Death; Cells; Cerebral cortex; Child; Coculture Techniques; Communication; Corpus striatum structure; Culture Techniques; Dendrites; Dendritic Spines; Development; Ethanol; Event; Exposure to; Fetal Alcohol Spectrum Disorder; Fetus; Fluorescent Dyes; Future; Genes; Genetic; Genotype; Glutamates; Growth; Hippocampus (Brain); Human; Image; Impairment; Infant; Knock-out; Lead; Learning Disorders; Length; Long-Term Effects; Measures; Mediating; Mediator of activation protein; Mental Depression; Mental Retardation; Messenger RNA; Mission; Modeling; Molecular; Morphology; Mus; N-Methylaspartate; National Institute on Alcohol Abuse and Alcoholism; Neonatal; Neonatal Alcohol Exposure; Neurons; Neurotrophic Tyrosine Kinase Receptor Type 2; Pathway interactions; Play; Predisposition; Pregnancy; Property; Proteins; Psychotic Disorders; Regulation; Research; Resistance; Rodent; Role; Severities; Signal Transduction; Structure; Synapses; Techniques; Testing; Therapeutic Intervention; Third Pregnancy Trimester; Time; Training; Twin Studies; Vertebral column; Wild Type Mouse; Work; adenylyl cyclase 1; alcohol effect; alcohol exposure; alcohol response; alcohol sensitivity; aspartate receptor; cognitive function; density; design; fetal; immunoreactivity; in utero; innovation; neonatal exposure; neuron development; neuron loss; neuronal cell body; neurotoxicity; postsynaptic; presynaptic; pup; ranpirnase; receptor expression; response; sedative; success; synaptogenesis; therapeutic target; therapy design

DESCRIPTION (provided by applicant): The overall objective of this proposal is to define the mechanisms underlying the deleterious effects caused by ethanol on neuronal morphology in a model of Fetal Alcohol Spectrum Disorder (FASD), thereby upholding the mission of the NIAAA to increase the understanding of biological functions relating to alcohol use. The synaptogenesis (brain growth spurt) period occurs during the last trimester of human gestation and is a critical window for dendritic growth and spine development in the developing brain. Dendritic spines are the primary structures used in synaptic communication between neurons. Exposure to ethanol during this period has been shown to have detrimental effects on the developing fetal brain, leading to learning disorders, behavioral abnormalities, and mental retardation, termed FASD. BDNF has been shown to regulate dendritic growth and spine development through its interaction with the trkB receptor. BDNF expression has also been shown to be modulated by ethanol in various regions of the neonatal brain. Fetal genotype has been shown to affect the severity of FASD as shown by twin studies, although the gene(s) involved have yet to be identified. Since studies have shown adult mice lacking the adenylyl cyclases 1 and 8 have an increased susceptibility to the sedative effects of ethanol, and the striatum of neonatal mice lacking these ACs are more sensitive to EtOH induced neuronal cell death than WT mice, we believe the ACs could play a role in the sequelae of neonatal EtOH exposure. This proposal will test the hypothesis that the Ca2+stimulated adenylyl cyclases (AC1 and AC8) regulate the dendritic response to ethanol through regulation of BDNF signaling. This hypothesis will be tested using the following Aims: 1) Determine the effects of ethanol exposure on neuronal morphology in the developing striatum from wild type (WT) and Ca2+stimulated AC knockout (DKO) mice; 2) Dissociate the pre- and postsynaptic roles of the Ca2+stimulated ACs in the regulation of dendritic complexity and spine formation; 3) Determine the effects of the Ca2+stimulated ACs on BDNF regulation in the neonatal striatum following ethanol exposure. In Aim 1, P5-7 pups (WT and DKO) will be treated with ethanol and striatal neurons will be analyzed at P14 and P30 for soma size, dendritic length, dendritic branching, and spine density. In Aim 2, corticostriatal co-cultures from WT and DKO mice will be used to determine the pre- and postsynaptic roles of the Ca2+stimulated ACs in ethanol-induced effects on dendritic arborization and spine formation. In Aim 3, BDNF mRNA and protein as well as trkB protein will be measured in synaptosomal fractions from ethanol-treated neonatal mice (P5-7) and confocal imaging of striatal neurons will be used to identify the pre- and postsynaptic roles of the Ca2+stimulated ACs in regulating BDNF expression. Training in the use of four main areas: primary neuronal culture techniques (corticostriatal co-cultures), immunohistochemical techniques/confocal analyses, use of diolistic fluorescent dye imaging and neuronal morphology analyses represents exciting opportunities of research for both the sponsor and applicant, greatly enhancing the applicant's potential for future scientific success. PUBLIC HEALTH RELEVANCE: The third trimester of gestation in humans is a critical time for brain development. Consumption of alcohol during this time can result in fetal neurotoxicity leading to behavioral abnormalities, learning disorders, mental retardation, depression, and psychosis termed fetal alcohol spectrum disorder (FASD). This proposal is designed to study the role of the adenylyl cyclases in the regulation of dendritic growth, spine development, and BDNF expression after neonatal alcohol exposure, which may lead to identification of therapeutic targets for children with FASD.

描述（由申请人提供）：本提案的总体目标是确定乙醇对胎儿酒精谱系障碍 (FASD) 模型中神经元形态造成有害影响的机制，从而维护 NIAAA 的使命，即提高胎儿酒精谱系障碍 (FASD) 模型中的神经元形态。了解与饮酒相关的生物功能。突触发生（大脑生长突增）期发生在人类妊娠的最后三个月，是发育中的大脑中树突生长和脊柱发育的关键窗口。树突棘是神经元之间突触通讯的主要结构。研究表明，在此期间接触乙醇会对发育中的胎儿大脑产生不利影响，导致学习障碍、行为异常和精神发育迟滞（称为 FASD）。 BDNF 已被证明可以通过与 trkB 受体的相互作用来调节树突生长和脊柱发育。新生儿大脑各个区域的 BDNF 表达也被证明受到乙醇的调节。 正如双胞胎研究所示，胎儿基因型已被证明会影响 FASD 的严重程度，但所涉及的基因尚未确定。由于研究表明缺乏腺苷酸环化酶 1 和 8 的成年小鼠对乙醇的镇静作用的敏感性增加，并且缺乏这些 AC 的新生小鼠的纹状体比 WT 小鼠对乙醇诱导的神经元细胞死亡更敏感，因此我们认为 AC可能在新生儿乙醇暴露的后遗症中发挥作用。该提案将检验 Ca2+ 刺激的腺苷酸环化酶（AC1 和 AC8）通过调节 BDNF 信号传导来调节树突对乙醇的反应的假设。 该假设将通过以下目标进行测试： 1) 确定乙醇暴露对野生型 (WT) 和 Ca2+刺激 AC 敲除 (DKO) 小鼠发育中纹状体神经元形态的影响； 2) 分离Ca2+刺激的ACs在树突复杂性和树突棘形成调节中的突触前和突触后作用； 3) 确定乙醇暴露后 Ca2+ 刺激的 AC 对新生儿纹状体 BDNF 调节的影响。 在目标 1 中，P5-7 幼仔（WT 和 DKO）将接受乙醇处理，并在 P14 和 P30 时分析纹状体神经元的体细胞大小、树突长度、树突分支和脊柱密度。在目标 2 中，来自 WT 和 DKO 小鼠的皮质纹状体共培养物将用于确定 Ca2+ 刺激的 AC 在乙醇诱导的树突分枝和脊柱形成作用中的突触前和突触后作用。在目标 3 中，将在乙醇处理的新生小鼠 (P5-7) 的突触体部分中测量 BDNF mRNA 和蛋白以及 trkB 蛋白，并使用纹状体神经元的共聚焦成像来识别 Ca2+ 的突触前和突触后作用。 +刺激 AC 调节 BDNF 表达。 四个主要领域的使用培训：原代神经元培养技术（皮质纹状体共培养）、免疫组织化学技术/共聚焦分析、二醇荧光染料成像和神经元形态分析的使用为申办者和申请人提供了令人兴奋的研究机会，极大地增强了研究能力申请人未来科学成功的潜力。 公众健康相关性：人类妊娠晚期是大脑发育的关键时期。在此期间饮酒可能会导致胎儿神经毒性，导致行为异常、学习障碍、智力低下、抑郁和精神病，称为胎儿酒精谱系障碍 (FASD)。该提案旨在研究腺苷酸环化酶在新生儿酒精暴露后调节树突生长、脊柱发育和 BDNF 表达中的作用，这可能有助于确定 FASD 儿童的治疗靶点。