快速眼动睡眠剥夺后大鼠皮质及海马神经元突触相关蛋白神经颗粒素表达的变化  

Rapid eye movement sleep deprivation regulates the expression of synaptic related protein neurogranin in rat cortex and hippocampus

作　　者：方明[1,2] 田国红[1] 黄流清[1] 周晖[1] 赵忠新[1] 

机构地区：[1]上海第二军医大学附属长征医院神经内科,上海200003 [2]湖北宜昌市第一人民医院神经内科

出　　处：《内科理论与实践》2007年第5期324-327,共4页Journal of Internal Medicine Concepts & Practice

基　　金：国家自然科学基金资助项目(项目编号:30270487);第二军医大学附属长征医院"十一五""三重三优"学科人才建设计划基金(项目编号:2005312)

摘　　要：目的:探讨不同时间的快速眼动(REM)睡眠剥夺对大鼠皮质及海马各区神经颗粒素分子表达变化的影响及意义。方法:Sprague-Dawley大鼠70只,随机分为睡眠剥夺组(SD)、实验环境对照组(TC)和空白对照组(CC)。其中SD组又分为12h、1d、3d、5d、7d共5个时点组。采用改良多平台(MMPM)睡眠剥夺法进行REM睡眠剥夺,运用免疫荧光染色共聚焦显微镜观察REM睡眠剥夺后不同时点大鼠皮质及海马各区的神经颗粒素表达的分布规律和时空变化;同时结合蛋白印迹(Western blot)技术对皮质及全海马神经颗粒素蛋白作选择性半定量分析。结果:神经颗粒素主要分布于正常大脑皮质Ⅱ、Ⅲ层的神经元胞体和树突、海马CA1～CA3锥体细胞层和齿状回颗粒细胞层内。REM睡眠剥夺12h后大鼠皮质神经颗粒素的表达即开始减少,与对照组比较有显著性差异,直至第7d均呈下降趋势;海马结构中神经颗粒素表达未见显著变化。蛋白印迹实验印证了这一结果。结论:REM睡眠剥夺能引起大鼠大脑皮质神经颗粒素表达减少,且随睡眠剥夺时间的延长而渐趋明显,这可能是REM睡眠剥夺引起大脑神经元突触可塑性改变,进而影响大鼠学习记忆功能损害的机制之一。Objective To explore the special and temporal patterns of neurogranin expression in the rat neocortical and hippocampal regions on different stages of rapid eye movement （REM） sleep deprivation. Methods An modified multiple platform methods （MMPM） were used to induce REM sleep deprivation. A total of 70 Sprague-Dawley rats were enrolled and divided randomly into following three groups： cage control （CC） group, tank control （TC） group and sleep deprivation （SD） group. The animals in SD group were sacrificed at the following time-points after SD： 12h, ld, 3d, 5d and 7d, respectively. By fluorescent immunostain combined with confocal microscopic examination, the expression and distribution of neurogranin, a special neuronal protein which contribute to the synaptic plasticity, were detected in the cerebral cortex and different regions of hippocampal. Western blot was used to analysis the level of protein expression, which involved in these related areas. Results Strong expression of neurogranin was demonstrated in cytoplasm of layers Ⅱ and Ⅲ cortical, CA1 to CA3 hippocampus pyramidal cells and dentate gyrus of the control groups. Rats after 12 h SD exhibited down-regulated neurogranin expression in the cortex significantly. With prolonged deprivation time, the density of immuno-positive cells reduced continuously until 7 d after SD. A similar trend was also demonstrated by the results of Western blot. Conclusions REM sleep deprivation might modulate the expression of neurogranin preferentially in cerebral cortex from the early beginning to nearly one week after SD. The result suggests that neurogranin might be one of neurobiological alterations leading to brain dysfunction such as learn and memory disturbance.

关 键 词：快速眼动睡眠剥夺 神经颗粒素 大脑皮质 海马 突触可塑性 

分 类 号：R740[医药卫生—神经病学与精神病学]