SLEEP AND SYNAPTIC PLASTICITY

关于睡眠功能的几种理论认为，睡眠起着维持、修复或巩固突触的作用。有几条证据支持这些理论，但到目前为止，还没有对这些假设进行直接测试。一份独立的大型文献将长时记忆任务与突触可塑性联系起来。此外，一些研究表明，睡眠对记忆巩固有明显影响，学习任务对睡眠有明显影响。然而，到目前为止，还没有关于睡眠对突触可塑性影响的研究，尽管这类研究已经有了理论基础，而且现在已经有了必要的分子工具和小鼠模型。为了检验睡眠在突触可塑性中的作用这一普遍假设，我们将使用非认知刺激(单侧胡须切断)和认知刺激(空间学习测试)分别在躯体感觉皮层和海马区诱导突触可塑性。在这些实验范例中，睡眠不足和过度睡眠对突触可塑性的各种分子标志物的变化幅度和速度的影响将被确定。待分析的标志物代表突触重塑所需的六类分子：细胞外基质蛋白、细胞黏附分子、生长因子、神经传递相关蛋白、细胞骨架及相关蛋白和转录因子。三个品系的小鼠将被使用，AKR/J，无基因小鼠和携带lac-Z基因的转基因品系依赖于核因子Kappa B的存在而偶联到启动子。除了量化大脑小区域提取液中突触可塑性的分子标记物的变化，我们还将使用组织化学、免疫细胞化学和原位杂交组织化学来确定这些分子在睡眠操作中反应的细胞类型的特定位置。预期结果将为睡眠功能的实验验证提供依据。

Several theories concerning sleep function posit that sleep serves to maintain, repair or consolidate synapses. Several lines of evidence support these theories but, to date, there has been no direct test of these hypotheses. An independent large literature relates long-term memory tasks to synaptic plasticity. Further, a few studies have demonstrated a clear influence of sleep on memory consolidation and of learning tasks on sleep. However, there are heretofore, no studies of the influence of sleep on synaptic plasticity despite the theoretical foundation for such studies and the fact that the necessary molecular tools and mouse models are now available. To test the general hypothesis that sleep has a role in synaptic plasticity we will use a non-cognitive stimulus (a unilateral whisker cut) and a cognitive stimulus (a spatial learning test) to induce synaptic plasticity in the somatosensory cotex and the hippocampus respectively. The effects of sleep loss and excess sleep in these experimental paradigms on the magnitude and rates of change of a variety of molecular markers of synaptic plasticity will be determined. The markers to be analyzed represent six classes of molecules necessary for synaptic remodeling; extracellular matrix proteins, cell adhesion molecules, growth factors, neurotransmission related proteins, cytoskeletal and associated proteins and transcription factors. Three strains of mice will be used, AKR/J, acallosal mice and a transgenic strain carrying the lac-Z gene coupled to a promoter dependent upon the presence of nuclear factor Kappa B. In addition to quantifying the changes in the molecular markers of synaptic plasticity in extracts of small areas of the brain, we will also determine the specific location of cell types in which these molecules change in response to sleep manipulation using histochemistry, immunocytochemistry and in situ hybridization histochemistry. Expected results will provide the foundations for the experimental verification of sleep function.