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

基本信息

批准号：
8520119
负责人：
Laura Susick
金额：
$ 5.39万
依托单位：
WAYNE STATE UNIVERSITY
依托单位国家：
美国
项目类别：
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/8520119
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 abstracting 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

项目摘要

Project Summary/Abstract 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 Ca[2+]-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 Ca[2+]-stimulated AC knockout (DKO) mice; 2) Dissociate the pre- and postsynaptic roles of the Ca[2+]-stimulated ACs in the regulation of dendritic complexity and spine formation; 3) Determine the effects of the Ca[2+]-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 Ca[2+]-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 Ca[2+]-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.

项目概要/摘要该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，增进对与酒精相关的生物功能的了解突触发生（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。神经元之间的突触通讯已被证明会影响胎儿大脑的发育，导致学习障碍、行为异常和精神发育迟滞（称为 FASD），它可通过调节树突生长和脊柱发育。它与 trkB 受体的相互作用也被证明可以在新生儿大脑的各个区域受到乙醇的调节。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，但相关基因尚未确定，因为研究表明缺乏腺苷酸环化酶 1 和 8 的成年小鼠对镇静剂的敏感性增加。乙醇的影响，以及缺乏这些 AC 的新生小鼠的纹状体对乙醇诱导的神经细胞死亡比 WT 小鼠更敏感，我们相信 AC 可能在乙醇的后遗症中发挥作用。该提案将检验 Ca[2+] 刺激的腺苷酸环化酶（AC1 和 AC8）通过调节 BDNF 信号传导来调节对乙醇的树突反应的假设。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 Ca[2+] 刺激的 AC 敲除 (DKO) 小鼠发育中的纹状体神经元形态的影响 2) 分离Ca[2+] 刺激的 AC 在树突复杂性和脊柱形成的调节中的突触前和突触后作用 3) 确定Ca[2+] 刺激 ACs 对乙醇暴露后新生儿纹状体 BDNF 的调节。在目标 1 中，P5-7 幼崽（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大小、树突长度、树突分支和脊柱密度。在目标 2 中，皮质纹状体共培养。来自 WT 和 DKO 小鼠的细胞将用于确定 Ca[2+] 刺激的 AC 在乙醇诱导的影响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 和蛋白质以及 trkB 蛋白质，并使用纹状体神经元的共聚焦成像来识别预-树突分枝和脊柱形成。 Ca[2+] 刺激的 ACs 在调节 BDNF 表达中的突触后作用。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。