Role of Microglial Fractalkine Signaling in Altered Dopaminergic Wiring in FASD

小胶质细胞分形蛋白信号传导在 FASD 多巴胺能线路改变中的作用

基本信息

批准号：
10666254
负责人：
Carlita Favero
金额：
$ 34.97万
依托单位：
URSINUS COLLEGE
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-08-15 至 2025-07-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10666254
关键词：
3-Dimensional Age Air Alcohol consumption Alcohols Area Astrocytes Attentional deficit Awareness Axon Behavioral Blood Body Size Brain CCL2 gene CX3CL1 gene Cells Childhood Cognitive Computer software Cues Data Development Dopamine Dysmorphology Embryo Ethanol Executive Dysfunction Face Fetal Alcohol Exposure Fetal Alcohol Spectrum Disorder Fractalkine Future Glial Fibrillary Acidic Protein Growth Growth Cones Hour Human Impairment Incidence Individual Inflammatory Inhalation Interleukin-1 beta Interleukin-6 Knock-out Knowledge Length Location Measures Mediating Messenger RNA Methods Microglia Modeling Molecular Morphology Mus Nervous System Neurobiology Neuroimmune Neurons Phagocytosis Play Pregnant Women Process Proteins Regulation Reporting Risk Role Signal Pathway Signal Transduction Substance Use Disorder TNF gene Technology Testing Three-dimensional analysis Time Transgenic Mice Tyrosine 3-Monooxygenase Visualization Western Blotting Work alcohol effect alcohol exposure axon growth axon guidance behavioral outcome binge drinking chemokine child bearing cytokine density dopaminergic neuron experimental study fractalkine receptor glial activation in vivo mRNA Expression male molecular marker mouse model multiplex assay neuroinflammation neutralizing antibody offspring pre-clinical pregnant protein expression receptor reconstruction response sex transmission process vapor white matter

项目摘要

PROJECT SUMMARY/ABSTRACT Despite public awareness campaigns, Fetal Alcohol Spectrum Disorders (FASD) remain prevalent due to alcohol consumption by women that are pregnant or of child-bearing age. Prenatal alcohol exposure disrupts (1) dopaminergic mesocorticolimbic projections which is likely related to the executive dysfunction, attention deficits, and increased risk for substance use disorders that characterize FASD, (2) the subpallium, a critical intermediate target during axon formation, and (3) guidance cues and signaling pathways that drive axon formation. In this study, we test the hypothesis that binge prenatal alcohol exposure hinders dopaminergic axon outgrowth in the subpallium through reduced signaling between dopamine axons and microglial cells via fractalkine. Our experiments aim to (1) define the developmental timing of alcohol-induced alterations in fractalkine signaling and (2) determine the role of these alterations in microglial number, activation, and regulation of dopaminergic axon guidance. To determine the developmental timing of alcohol effects on fractalkine signaling, we will measure in vivo levels of mRNA and protein fractalkine (CX3CL1), its receptor (CX3CR1), and resultant cytokines (IL-1β, TNFα, IL-6, MIP-1α, and MCP-1) before, during, and after microglial guidance of dopaminergic axons. To visualize alcohol effects on microglial activation in the subpallium, we will count immunostained microglia and perform 3D reconstructions to analyze morphology using Imaris and Halo technologies. To measure dopaminergic axon outgrowth, we will trace tyrosine hydroxylase (TH) immunostained axons. To confirm the requirement of fractalkine signaling in these effects, we will utilize CX3CR1eGFP/eGFP mice which serve as functional knockouts of the fractalkine receptor. Our anticipated findings will reveal the impact of developmental alcohol exposure on cellular and molecular mechanisms that are required for proper nervous system wiring, an area that is currently understudied. This work will enhance understanding of neurobiological underpinnings of neuroinflammation and dampened dopamine function that are observed in human FASD.

项目概要/摘要尽管开展了公众宣传活动，胎儿酒精谱系障碍 (FASD) 仍然存在由于怀孕或生育妇女饮酒而流行产前酒精暴露会扰乱 (1) 多巴胺能中皮质边缘投射。这可能与执行功能障碍、注意力缺陷和风险增加有关对于以 FASD 为特征的物质使用障碍，(2) 大脑皮层下，一个关键的轴突形成过程中的中间目标，以及（3）引导线索和信号通路在这项研究中，我们检验了产前酗酒的假设。暴露通过减少大脑皮层中的多巴胺能轴突生长我们的实验通过 fractalkine 实现多巴胺轴突和小胶质细胞之间的信号传导。目的是 (1) 确定酒精引起的 fractalkine 变化的发育时间信号传导和（2）确定这些改变在小胶质细胞数量、激活、和多巴胺能轴突引导的调节以确定发育时间。为了了解酒精对 fractalkine 信号传导的影响，我们将测量 mRNA 的体内水平，蛋白质 fractalkine (CX3CL1)、其受体 (CX3CR1) 以及由此产生的细胞因子 (IL-1β、 TNFα、IL-6、MIP-1α 和 MCP-1）在小胶质细胞引导之前、期间和之后观察酒精对多巴胺能轴突的影响。大脑皮层下，我们将计数免疫染色的小胶质细胞并进行 3D 重建使用 Imaris 和 Halo 技术分析形态以测量多巴胺能轴突。生长，我们将追踪酪氨酸羟化酶（TH）免疫染色的轴突以确认。在这些效应中需要 fractalkine 信号传导，我们将利用 CX3CR1eGFP/eGFP 小鼠我们预期的发现将作为 fractalkine 受体的功能性敲除。揭示发育时期酒精暴露对细胞和分子的影响适当的神经系统接线所需的机制，这是目前正在研究的领域这项工作将增强对神经生物学基础的理解。在人类 FASD 中观察到的神经炎症和多巴胺功能减弱。