Connectivity of Adult-Generated Dentate Granule Cells in a Mouse Model of Prenatal Alcohol Exposure

项目摘要/摘要 这项拟议的研究的总体目标是描述成年人网络连接的变化- 在产前酒精暴露(PAE)的小鼠模型中产生齿状颗粒细胞(ADGC)。如果成功， 这些研究可能揭示与胎儿酒精有关的海马体功能缺陷的机制。 谱系障碍(FASD)。目前的提议旨在检验PAE扰乱 成年海马齿状回神经元连接性的经验依赖性重塑。 这项提议的前提是之前的工作表明，即使是适度的酒精暴露也是如此 在整个妊娠期间，对丰富的环境和改变的突触会导致神经性反应受损 电生理学检测aDGC的活性。我将利用一个具有良好特性的有限访问妊娠期 建立小鼠暴露模型，研究PAE对aDGC电路连接性的影响。具体来说，我会 利用双矢量跟踪、高分辨率共聚焦成像、电生理和 解决以下目标的光遗传学方法。具体目标1.检验PAE的假设 阻断EE介导的传入突触传入aDGC的重塑。在这里，我们将评估 使用双向量追踪系统对aDGC的单突触传入输入分布的PAE(目标1.1)， ADC树突状细胞复杂性和棘突成熟的形态分析(AIM 1.2)和功能性突触 使用诱发突触活动的全细胞贴片记录输入aDGC(目标1.3)。具体目标2. 验证PAE干扰EE介导的传出突触输出重塑的假设 ADGC。在这里，我将评估PAE对aDGC与CA3锥体兴奋性传出联系的影响 使用神经解剖学方法(目标2.1)的细胞和中间神经元，结合光遗传刺激 海马片制备中的aDGCs(目标2.2)。这些目标与NIAAA实现的使命不谋而合 改善酒精相关问题的基础知识，因为了解PAE如何影响 海马区的连接可能揭示基于电路的新治疗方法的机会，以缓解 临床FASD的神经行为后果。重要的是，拟议的研究提供了一个框架， 酒精相关神经科学的研究培训使用最先进的方法在杰出的 新墨西哥州酒精研究中心内的指导团队和研究环境。

PROJECT SUMMARY/ABSTRACT The overall goal of the proposed research is to characterize alterations in network connectivity of adult- generated dentate granule cells (aDGCs) in a mouse model of prenatal alcohol exposure (PAE). If successful, these studies may reveal mechanisms that underlie deficits in hippocampal function associated with fetal alcohol spectrum disorders (FASDs). The current proposal is designed to test the overall hypothesis that PAE disrupts experience-dependent remodeling of connectivity of newborn dentate granule neurons in the adult hippocampus. The proposal is premised on previous work demonstrating that exposure to even moderate levels of alcohol throughout gestation leads to an impaired neurogenic response to enriched environment and altered synaptic activity of aDGCs as assessed electrophysiologically. I will utilize a well-characterized limited access gestational exposure model in mice to characterize the impact of PAE on circuit connectivity of aDGCs. Specifically, I will utilize a combination of dual-vector tracing, high-resolution confocal imaging, electrophysiological and optogenetic approaches to address the following aims. Specific Aim 1. To test the hypothesis that PAE disrupts EE-mediated remodeling of afferent synaptic input to aDGCs. Here we will assess the impact of PAE on the distribution of monosynaptic afferent input to aDGCs using a dual vector tracing system (Aim 1.1), morphological analysis of dendritic complexity and spine maturation in aDGCs (Aim 1.2), and functional synaptic input to aDGCs using whole-cell patch slice recordings of evoked synaptic activity (Aim 1.3). Specific Aim 2. To test the hypothesis that PAE disrupts EE-mediated remodeling of efferent synaptic output from aDGCs. Here, I will assess the impact of PAE on excitatory efferent connections of aDGCs with CA3 pyramidal cells and interneurons using a neuroanatomical approach (Aim 2.1), coupled with optogenetic stimulation of aDGCs in hippocampal slice preparations (Aim 2.2). These aims coincide with the mission of the NIAAA to attain fundamental knowledge for the improvement of alcohol-related problems, since understanding how PAE impacts hippocampal connectivity may reveal opportunities for novel circuitry-based therapeutic approaches to mitigate neurobehavioral consequences in clinical FASDs. Importantly, the proposed research provides a framework for research training in alcohol-related neuroscience utilizing state-of-the art approaches within an outstanding mentoring team and research environment within the New Mexico Alcohol Research Center.