The transcription and translation of the immediate-early genes(IEGs),such as Arc,Zif268,eFos and Npas4,are considered to be molecular mechanisms underlying learning and memory.Recent studies have shown that applicatio...The transcription and translation of the immediate-early genes(IEGs),such as Arc,Zif268,eFos and Npas4,are considered to be molecular mechanisms underlying learning and memory.Recent studies have shown that application of NADH triggered the expression of Arc and Zif268 by activation of NMDA receptor(NMDAR)signaling.In this regard,we hypothesize that levels of Npas4 and c Fos,could be modulated by NMDAR activity-dependent NADH.To test our hypothesis,in primary cultured hippocampal neurons,we employed NADH to examine the effects of NADH on IEGs.Further more,NMDARs blocker MK801 were used to investigated the role of NMDAR in NADH's effects.The data of real-time PCR revealed that application of NADH promoted the transcription of Arc,cFos and Npas4.Similarly,Western blot analyses showed that the levels of Arc,c Fos and Npas4 were increased significantly after NADH treatment.展开更多
Circular RNA is a type of non-coding RNA which forms a loop structure.CircRNA was reported to exhibit cell-and tissue-specific patterns.Although accumulating evidences have showed circRNA plays an important role in ne...Circular RNA is a type of non-coding RNA which forms a loop structure.CircRNA was reported to exhibit cell-and tissue-specific patterns.Although accumulating evidences have showed circRNA plays an important role in neural development and neuropsychiatric diseases,there is lttle knowledge between circRNA and methamphetamine(MA).Using primary neuron culture,we identified the differential circRNAs with MA treatment through RNA sequencing.After RT-PCR analysis,circTLk1 expression was significant reduced.Moreover,we produced MA addicted mouse models with conditioned place preference test.The different brain area including hippocampus,prefrontal cortex,ventral tegmental area(VTA)and nucleus accubens were isolated and determined circTLk1's expression.展开更多
目的研究中缝背核(dorsal raphe nucleus,DRN)在甲基苯丙胺(methamphetamine,METH)成瘾中的作用。方法将C57BL/6J小鼠随机分为METH模型组和对照组。通过腹腔注射METH(2mg/kg)制备条件性位置偏爱(conditioned place preference,CPP)动物...目的研究中缝背核(dorsal raphe nucleus,DRN)在甲基苯丙胺(methamphetamine,METH)成瘾中的作用。方法将C57BL/6J小鼠随机分为METH模型组和对照组。通过腹腔注射METH(2mg/kg)制备条件性位置偏爱(conditioned place preference,CPP)动物模型,对照组注射等量生理盐水。经即刻早基因c-Fos免疫组织化学染色、含Ca^(2+)指示剂病毒载体脑内注射、化学遗传学和蛋白质谱分析等技术,探究METH激活DRN脑区情况及蛋白表达变化,并分析抑制DRN脑区神经元CPP评分的变化。结果CPP评分显示,METH模型小鼠在伴药箱产生明显偏好(t=4.438,P<0.05)。METH模型组DRN内c-Fos阳性细胞数较对照组显著增多(t=4.065,P<0.05),而且DRN神经元内Ca^(2+)荧光信号明显增强(t=20.50,P<0.001)。通过叠氮平-N-氧化物(clozapine-N-oxide,CNO)腹腔注射(3mg/kg)诱导hM4Di表达抑制DRN神经元兴奋性,明显降低了METH模型组小鼠CPP分数(t=3.26,P<0.05)。蛋白质谱分析在METH模型组DRN脑区筛出了一批差异蛋白,其中突触囊泡蛋白1(Syt1)和细胞黏附分子1(Cadm1)表达升高(均P<0.05);神经束蛋白(Nfasc)和突触小泡蛋白2(SV2)的C亚型表达降低(均P<0.05)。结论DRN脑区在METH诱导小鼠CPP行为偏好中起着重要作用,METH特异性诱导DRN内与突触相关的蛋白分子的表达,可能与DRN参与的成瘾机制有关。展开更多
文摘The transcription and translation of the immediate-early genes(IEGs),such as Arc,Zif268,eFos and Npas4,are considered to be molecular mechanisms underlying learning and memory.Recent studies have shown that application of NADH triggered the expression of Arc and Zif268 by activation of NMDA receptor(NMDAR)signaling.In this regard,we hypothesize that levels of Npas4 and c Fos,could be modulated by NMDAR activity-dependent NADH.To test our hypothesis,in primary cultured hippocampal neurons,we employed NADH to examine the effects of NADH on IEGs.Further more,NMDARs blocker MK801 were used to investigated the role of NMDAR in NADH's effects.The data of real-time PCR revealed that application of NADH promoted the transcription of Arc,cFos and Npas4.Similarly,Western blot analyses showed that the levels of Arc,c Fos and Npas4 were increased significantly after NADH treatment.
文摘Circular RNA is a type of non-coding RNA which forms a loop structure.CircRNA was reported to exhibit cell-and tissue-specific patterns.Although accumulating evidences have showed circRNA plays an important role in neural development and neuropsychiatric diseases,there is lttle knowledge between circRNA and methamphetamine(MA).Using primary neuron culture,we identified the differential circRNAs with MA treatment through RNA sequencing.After RT-PCR analysis,circTLk1 expression was significant reduced.Moreover,we produced MA addicted mouse models with conditioned place preference test.The different brain area including hippocampus,prefrontal cortex,ventral tegmental area(VTA)and nucleus accubens were isolated and determined circTLk1's expression.
文摘目的研究中缝背核(dorsal raphe nucleus,DRN)在甲基苯丙胺(methamphetamine,METH)成瘾中的作用。方法将C57BL/6J小鼠随机分为METH模型组和对照组。通过腹腔注射METH(2mg/kg)制备条件性位置偏爱(conditioned place preference,CPP)动物模型,对照组注射等量生理盐水。经即刻早基因c-Fos免疫组织化学染色、含Ca^(2+)指示剂病毒载体脑内注射、化学遗传学和蛋白质谱分析等技术,探究METH激活DRN脑区情况及蛋白表达变化,并分析抑制DRN脑区神经元CPP评分的变化。结果CPP评分显示,METH模型小鼠在伴药箱产生明显偏好(t=4.438,P<0.05)。METH模型组DRN内c-Fos阳性细胞数较对照组显著增多(t=4.065,P<0.05),而且DRN神经元内Ca^(2+)荧光信号明显增强(t=20.50,P<0.001)。通过叠氮平-N-氧化物(clozapine-N-oxide,CNO)腹腔注射(3mg/kg)诱导hM4Di表达抑制DRN神经元兴奋性,明显降低了METH模型组小鼠CPP分数(t=3.26,P<0.05)。蛋白质谱分析在METH模型组DRN脑区筛出了一批差异蛋白,其中突触囊泡蛋白1(Syt1)和细胞黏附分子1(Cadm1)表达升高(均P<0.05);神经束蛋白(Nfasc)和突触小泡蛋白2(SV2)的C亚型表达降低(均P<0.05)。结论DRN脑区在METH诱导小鼠CPP行为偏好中起着重要作用,METH特异性诱导DRN内与突触相关的蛋白分子的表达,可能与DRN参与的成瘾机制有关。
