SLEEP AND SYNAPTIC PLASTICITY

一些关于睡眠功能的理论认为睡眠有助于维持、修复或巩固突触。有几条证据支持这些理论，但迄今为止，还没有对这些假设进行直接测试。一个独立的大型文献将长期记忆任务与突触可塑性联系起来。此外，一些研究已经证明了睡眠对记忆巩固和学习任务对睡眠的明显影响。然而，迄今为止，尽管研究的理论基础和必要的分子工具和小鼠模型已经具备，但还没有关于睡眠对突触可塑性影响的研究。为了验证睡眠在突触可塑性中起作用的一般假设，我们将使用非认知刺激（单侧须切割）和认知刺激（空间学习测试）分别诱导体感皮层和海马的突触可塑性。在这些实验范式中，睡眠不足和睡眠过量对突触可塑性各种分子标记的变化幅度和速率的影响将被确定。待分析的标记物代表了突触重塑所必需的六类分子；细胞外基质蛋白、细胞粘附分子、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。将使用三种小鼠品系，AKR/J，非allosal小鼠和携带lacz基因的转基因品系，该基因与依赖于核因子Kappa b存在的启动子偶联，除了量化大脑小区域提取物中突触可塑性分子标记的变化外，我们还将确定细胞类型的特定位置，这些分子在响应睡眠操作时也会发生变化。免疫细胞化学和原位杂交组织化学。预期结果将为睡眠功能的实验验证提供基础。

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.