【目的】 观察幼鼠快速眼动(rapid eye movement,REM)睡眠剥夺及恢复过程中海马GAP-43表达水平,探讨睡眠影响学习记忆功能的突触前机制。 【方法】 将30只雄性Sprague-Dawle幼鼠随机分为5组:空白对照组、环境对照组、48 h REM睡眠剥夺组、睡眠恢复24 h组、睡眠恢复48 h组;每组6只。采用多平台改良法建立幼鼠REM睡眠剥夺及睡眠恢复模型。应用RT-PCR技术和Western-blot技术分别检测GAP-43基因和蛋白表达水平;并对幼鼠海马切片进行免疫荧光组化分析。 【结果】 48 h睡眠剥夺组Gap-43基因表达显著高于其他各组(P<0.05);该组GAP-43蛋白及其水解片断GAP-43-3在海马组织的表达量上升,但磷酸化程度下降;而睡眠恢复组的GAP-43蛋白及其水解片断GAP-43-3在海马组织的表达量下降,磷酸化程度有所上升; 同时,48 hREM睡眠剥夺组和恢复睡眠24 h组的幼鼠海马组织GAP-43分布范围扩大。 【结论】 REM睡眠于转录、翻译和翻译后修饰三个层次作用于GAP-43,影响其功能状态,从而实现对学习记忆功能的突触前调控作用。
Abstract
【Objective】 To investigate the change of GAP-43 expressed in rats hippocampus after rapid eye movement (REM) sleep deprivation and the sleep recovery,and further to explore the possible pre-synaptic mechanisms of learning and memory. 【Methods】 Male Sprague-Dawley rats(n=30)were divided randomly into 5 groups:blank control group,environment control group,48 h REM sleep deprivation group,sleep recovery group (24 h),and sleep recovery group (48 h).The REM sleep deprivation model was established by the modified multiple platform methods.The gene and protein expression of GAP-43 were detected by RT-PCR and Western-blot.Hippocampus slices in rats were observed by immunohistochemistry method. 【Results】 The expression of Gap-43 gene in 48 h sleep deprivation group was significantly higher than that of the other groups (P<0.05).The expression of GAP-43 protein and its hydrolysis fragment,GAP-43-3 were increased,and the expression of phosphorylation were decreased.But in sleep recovery groups,the expression of GAP-43 protein and GAP-43-3 were decreased,and the expression of phosphorylation were increased.At the same time,the distribution of GAP-43 in rats hippocampus were expanded in 48 h REM sleep deprivation group and 24 h recovery sleep group. 【Conclusions】 The level of GAP-43 expression in rats hippocampus are affected by REM sleep through three ways,transcription,translation and post-translational modification.And finally result in the function of presynaptic regulation in learning and memory.
关键词
REM睡眠剥夺 /
GAP-43 /
突触可塑性 /
大鼠
Key words
rapid eye movement sleep deprivation /
GAP-43 /
synaptic plasticity /
rat
{{custom_sec.title}}
{{custom_sec.title}}
{{custom_sec.content}}
参考文献
[1] Diekelmann S,Born J.The memory function of sleep[J].Nat Rev Neurosci,2010,11:114-26.
[2] Jessica D.Payne,learning,memory,and sleep in humans[J].Sleep Medicine Clinics,2011,6(1):15-30.
[3] Walker MP,Stiekgold R.Sleep-dependent learning and memory consolidation[J].Neuron,2004,44:121-133.
[4] Davis CJ,Harding JW,Wright JW,et al.REM sleep deprivation-induced deficits in the latency-to-peak induction and maintenance of long-term potentiation within the CAI region of the hippocampus[J].Brain Res,2003,973:293-297.
[5] Chen PE,Errington ML,Kneussel M,et al.Behavioral deficits and subregion-specific suppress ion of LTP in mice expressing a population of mutant NMDA receptors throughout the hippocampus[J].Learn Mem,2009,16(10):635-644.
[6] Basheer R,Brown R,Ramesh V,et al.Sleep deprivation-induced protein changes in basal forebrain:Implications for synaptic plasticity[J].Neurosci Res,2005,82:650-658.
[7] Benowitz LI,Routtenberg A.GAP-43:an intrinsic determinant of neuronal development and plasticity[J].Trends Neurosci,1997,20:84-91.
[8] Zakharov VV,Bogdanova MN,Mosevitsky MI.Specific proteolysis of neuronal protein GAP-43 by calpain: characterization,regulation,and physiological role[J].Biochemistry,2005,70:897-907.
[9] Frey U,Morris RGM.Synaptic tagging: implications for late maintenance of hippocampal long-term potentiation[J].Trends Neurosci,1998,21:181-188.
[10] 江帆,沈晓明,李生慧,等.睡眠剥夺对幼鼠学习记忆的影响及相关基因[J].中国行为医学科学,2008,17(11):972-975.
[11] Yang RH,Hou XH,Xu XN,et al.Sleep deprivation impairs spatial learning and modifies the hippocampal theta rhythm in rats[J].Neuroscience,2011,173:116-123.
[12] Burwell RD,Saddoris MP,Bucci DJ,et al.Corticohippocampal contributions to spatial and contextual learning[J].Neurosci,2004,24:3826-3836.
[13] 张守信,金连弘.神经生物学[M].北京:科学出版社,2002:474-529.
[14] Dudman JT,Say DT,Siegelbaum SA.A role for synaptic inputs at distal d end rites:instructive signals for hippocampal long-term plasticity[J].Neuron,2007,56:866-879.
[15] Santschi LA,Zhang XL,Stanton PK.Activation of receptors negatively coupled to adenylate cyclase is required for induction of long-term synaptic depression at chaffer collateral-CA1 synapses[J].Neurobio,2006,66:205-219.
基金
国家自然科学基金面上项目(81072314,30700670);上海市教委科研创新项目(13YZ034);2009年度上海交通大学医学院新百人计划;2012年度上海市公共卫生优秀学科带头人培养计划(GWDTR201222)
PDF(661 KB)
