Long-lasting consequences of early ethanol on network activity during sleep

摘要/总结 胎儿酒精谱系障碍（FASD）是西方国家智力残疾的主要原因之一， 有神经行为特征，如学习、记忆和情绪方面的缺陷。我们认为，发展 乙醇（EtOH）暴露可能会导致睡眠期间神经活动模式的长期中断， 已知对记忆巩固和突触稳态很重要。如果是这样的话， 其中神经系统在睡眠期间自我修复和重新调整的正常能力将受到损害， 导致在EtOH暴露结束后很长时间内对神经系统功能的日常损害。为支持这一 假设，在过去的资助周期中，我们已经证明，发展EtOH 1）破坏和 成人非快速眼动睡眠的片段，2）成人睡眠中断的程度预测认知障碍，3） 睡眠障碍与小白蛋白（PV）和生长抑素（SST）表达的严重缺失相关， 已知GABA能中间神经元有助于睡眠-觉醒结构，4）PV细胞的破坏 通过敲除神经正五聚蛋白的功能复制了乙醇发育的某些方面，但不是睡眠， 5)用LiCl防止发育中的EtOH诱导的细胞丢失会阻止睡眠、解剖和认知 损伤在这次更新中，我们计划进一步探索发育EtOH对睡眠影响的机制 醒来时发现有细胞特异性分析目的1将检验以下假设： 备用的GABA能神经元将恢复发育中EtOH诱导的睡眠和认知缺陷， 转基因小鼠（例如，SST-Cre;目标1.1）。在同样的动物中，我们将睡眠-觉醒 功能/功能障碍/修复与几个确定的GABA能回路的解剖结构（例如，SST，卷轴， 血管活性肠肽（VIP））在睡眠相关区域，包括基底前脑核，海马和 新皮层数据将包括使用共聚焦和电子显微镜鉴定的细胞类型的突触相互作用 以及高尔基体分析（Aim 1.2）。对我们检测过的动物进行解剖学分析 睡眠/觉醒和认知功能将允许我们直接关联结构（例如，突触密度）， 神经行为功能/功能障碍/修复（例如，慢波活动，兴奋/抑制[E/I]平衡）。目标2.1 检查PV/SST细胞在多大程度上被发育中的EtOH杀死，或者下调 通过追踪荧光标记的PV/SST神经元祖细胞在Nkx2.1-Cre; Ai9中的命运的表型表达 暴露于EtOH的小鼠。这将提供指导尝试恢复GABA能功能所需的信息 在EtOH暴露后。目的2.2将PV/SST中间神经元祖细胞移植到乙醇处理的小鼠体内， 如果在所识别的脑区域中重新填充GABA能神经元将恢复睡眠和/或皮层活动（例如，表示"慢"之义 波活动，E/I平衡），以及睡眠相关的认知功能和电路结构。如果成功，结果 从这个建议可以打开一个新的窗口，了解和修复早期乙醇诱导的神经元损伤。 和认知障碍。

ABSTRACT/SUMMARY Fetal alcohol spectrum disorder (FASD) is one of the primary causes of intellectual disability in western nations, with neurobehavioral hallmarks such as deficits in learning, memory and mood. We propose that developmental ethanol (EtOH) exposure may induce long-lasting disruption of neural activity patterns during sleep which are known to be important for memory consolidation and synaptic homeostasis. If so, this would create a situation wherein the normal ability of the nervous system to repair and readjust itself during sleep would be impaired, resulting in a daily insult to nervous system function long after the EtOH exposure ended. In support of this hypothesis, during the past funding cycle we have demonstrated that developmental EtOH 1) disrupts and fragments adult non-REM sleep, 2) the extent of sleep disruption in adults predicts cognitive impairment, 3) the sleep impairment is associated with severe loss of parvalbumin (PV) and somatostatin (SST) expressing GABAergic interneurons which are known to contribute to sleep-wake architecture, 4) disruption of PV cell function through neuropentraxin knock-out replicates some aspects of developmental ethanol but not sleep, and 5) preventing cell loss induced by developmental EtOH with LiCl prevents sleep, anatomical, and cognitive impairments. In this renewal, we plan to further explore the mechanisms of developmental EtOH effects on sleep and waking with cell specific analyses. Aim 1 will test the hypothesis that selective optogenetic activation of spared GABAergic neurons will restore the deficits in sleep and cognition induced by developmental EtOH in transgenic mice (e.g., SST-Cre; Aim 1.1). In the same animals, we will correlate sleep-wake function/dysfunction/repair with anatomical structure of several identified GABAergic circuits (e.g., SST, reelin, vasoactive intestinal peptide (VIP)) in sleep-related regions including basal forebrain nuclei, hippocampus and neocortex. Data will include synaptic interactions of identified cell types using confocal and electron microscopy as well as Golgi analyses (Aim 1.2). Performing the anatomical analyses on animals that we have assayed for sleep/wake and cognitive function will allow us to directly relate structure (e.g., synaptic density) with neurobehavioral function/dysfunction/repair (e.g., slow-wave activity, excitatory/inhibitory [E/I] balance). Aim 2.1 examines to what extent PV/SST cells are killed by developmental EtOH, or alternatively have down-regulated phenotypic expression by tracking the fate of fluorescently labeled PV/SST neuron progenitors in Nkx2.1-Cre;Ai9 mice exposed to EtOH. This will provide information needed to guide attempts to restore GABAergic function after EtOH exposure. Aim 2.2 will transplant PV/SST interneuron progenitors into EtOH treated mice, to examine if re-populating GABAergic neurons in identified brain regions will restore sleep and/or cortical activity (e.g., slow- wave activity, E/I balance), as well as sleep-related cognitive function and circuit structure. If successful, results from this proposal could open a new window into both understanding and repair of early ethanol-induced neural and cognitive impairment.