Impact of Prenatal Alcohol Exposure on Receptor Targeting and Functioning

描述（由申请人提供）：在人类和实验动物模型中，在发育期间暴露于酒精会对中枢神经系统的结构和功能产生多种剂量依赖性影响，导致一系列身体、行为、认知和社交功能障碍，统称为胎儿酒精谱系障碍（FASD）。n -甲基- d -天冬氨酸（NMDA）受体（NMDAR）已被证明在学习和记忆中起着关键作用，因此一直是研究的焦点，旨在确定产前酒精暴露（PAE）对认知的有害影响的机制。本研究旨在验证PAE改变NMDAR亚基在突触和突触外室之间的分布（aim 1）和/或NMDAR通道复合物的性质（aim 2）导致小鼠齿状回NMDAR功能受损的假设。为了验证这一假设，研究人员提出了两个特定的目的：目的1:PAE改变齿状回突触和突触外区间NMDAR的分布；目的1a：确定对照组和FASD小鼠在基础状态和激活状态下突触和突触外NMDAR亚基的水平。目的1b：评估PAE在基础和激活条件下对NMDAR亚基定位控制机制的影响。目的1b1：在对照和FASD小鼠中，确定NMDA受体亚基与突触支架蛋白（PSD-95、PSD-93和SAP-102）的关联。目的1b2：评估PAE对对照和FASD小鼠NMDAR亚基磷酸化和总（磷酸化+非磷酸化）形式水平的影响。目的2：齿状回颗粒细胞中NMDAR功能受损。目的2a：在穿孔通路刺激后，在对照和FASD小鼠制备的海马切片中测量全细胞突触NMDAR依赖电流。目的2b：测量对照组和FASD小鼠海马切片中全细胞nmda依赖性突触和突触外电流。

DESCRIPTION (provided by applicant): In both humans and laboratory animal models, exposure to alcohol during development has been shown to cause a multitude of dose-dependent effects on the structure and function of the central nervous system, resulting in a range of physical, behavioral, cognitive and social dysfunctions that are collectively termed fetal alcohol spectrum disorders (FASD). The N-methyl-D-aspartate (NMDA) receptor (NMDAR) has been shown to play a critical role in learning and memory, and consequently has been a focus of studies aiming to identify mechanisms that underlie the detrimental effects of prenatal alcohol exposure (PAE) on cognition. The proposed studies aim to test the hypothesis that PAE alters the distribution of NMDAR subunits between synaptic and extrasynaptic compartments (Aim 1) and/or the properties of the NMDAR-channel complex (Aim 2) leading to impaired NMDAR function in the mouse dentate gyrus. Two specific aims have been developed to test this hypothesis: Aim 1: PAE alters the distribution of NMDAR between synaptic and extrasynaptic compartments in the dentate gyrus Aim 1a: Determine the levels of synaptic and extrasynaptic NMDAR subunits in control and FASD mice under basal and activated states. Aim 1b: Assess the impact of PAE on mechanisms controlling the localization of NMDAR subunits under basal and activated conditions. Aim 1b1: Determine the associations of NMDA receptor subunits with synaptic scaffolding proteins (PSD-95, PSD-93, and SAP-102) in control and FASD mice. Aim 1b2: Assess the impact of PAE on the levels of phosphorylated and total (phosphorylated + non- phosphorylated) forms of NMDAR subunits in control and FASD mice. Aim 2: NMDAR function is impaired in the dentate gyrus granule cells Aim 2a: Measure whole-cell synaptic NMDAR-dependent currents in hippocampal slices prepared from control and FASD mice following perforant pathway stimulation. Aim 2b: Measure whole-cell NMDA-dependent synaptic and extrasynaptic currents in hippocampal slices prepared from control and FASD mice. PUBLIC HEALTH RELEVANCE: Exposure to alcohol during development has been shown to cause a multitude of dose-dependent effects on the structure and function of the central nervous system. These effects are manifested as a range of physical, behavioral, cognitive and social dysfunctions. The studies proposed in this grant aim to identify neurochemical mechanisms that underlie the damaging effects of prenatal alcohol exposure on cognition, and thus identify novel targets for therapeutic intervention in the treatment of fetal alcohol spectrum disorders (FASD).