目的探讨Arc基因表达及甲基化与幼龄鼠和成年鼠空间记忆形成的相关性。方法应用Morris水迷宫对2月龄幼龄鼠和6月龄成年鼠进行定位航行训练,以判断其空间记忆形成情况。采用RT-PCR检测定位航行训练后大鼠海马组织Arc基因mRNA的转录水平,...目的探讨Arc基因表达及甲基化与幼龄鼠和成年鼠空间记忆形成的相关性。方法应用Morris水迷宫对2月龄幼龄鼠和6月龄成年鼠进行定位航行训练,以判断其空间记忆形成情况。采用RT-PCR检测定位航行训练后大鼠海马组织Arc基因mRNA的转录水平,并对海马基因组DNA进行亚硫酸盐处理,采用甲基化特异性聚合酶链反应(Methylation specific polymerase chain reaction,MS-PCR)检测Arc基因的甲基化情况,并进行DNA甲基化序列分析。结果随着定位航行训练入水次序及训练天数的增加,幼龄鼠逃避潜伏期缩短(P<0.05);而成年鼠在训练当天随着入水次序的增加,逃避潜伏期缩短,但第2天逃避潜伏期不仅与入水次序有关,也随着放入点距离的增大,逃避潜伏期相对延长(P<0.05),至第3天逃避潜伏期与入水次序及距离均无关,形成稳定的记忆能力。成年鼠定位航行训练组海马组织Arc基因mRNA的转录水平显著高于正常对照组(P<0.01),且高于幼龄鼠(P<0.05)。与幼龄鼠正常对照组相比,幼龄鼠和成年鼠定位航行训练组第8和24位点均无甲基化,成年鼠定位航行训练组与正常对照组相比,甲基化位点无变化。结论幼龄鼠瞬时记忆能力接近成年鼠,但形成稳定记忆的能力弱于成年鼠。Arc启动子甲基化影响其mRNA的表达,而Arc基因mRNA的转录水平与大鼠空间记忆能力及年龄相关,进一步证实了甲基化参与调解学习和记忆。展开更多
Silicon （Si） incorporated porous TiO2 coating （Si-TiO2） prepared on titanium （Ti） by micro-arc oxidation （MAO） technique was demonstrated to be cytocompatible in previous studies. In view of the potential clin...Silicon （Si） incorporated porous TiO2 coating （Si-TiO2） prepared on titanium （Ti） by micro-arc oxidation （MAO） technique was demonstrated to be cytocompatible in previous studies. In view of the potential clinical applications, a detailed in vitro study of the biological activity of Si-Ti02 coating, in terms of osteoblast （MC3T3-EI cells） morphology, proliferation, differentiation and mineralization was performed. Immunofluo- rescent staining indicated that cells seeded on the Si-TiO2 coating showed improved adhesion with developing mature cytoskeletons, which contained numerous distinct and well-defined actin stress fibers in the cell mem- branes compared with those on the Ti02 coating and Ti plate. Results from proliferation assay showed that the proliferation rate of cells seeded on the Si-TiO2coating was significantly faster than that on the TiO2 coating and Ti plate. Furthermore, the analysis of osteogenic gene expression demonstrated that the Si-Ti02 coating stimulated the expression of osteoblast-related genes and promoted differentiation and mineralization of MC3T3-EI cells. In addition, the Si-TiO2 coating differentially regulated Wnt signaling pathway by up-regulating the expression of low-density lipoprotein （LDL） receptor-related protein 5 （LrpS）, and downregulating the expression of Dickkopf-1 （Dkkl）. All together, these results indicate that the investigated titanium with Si-TiO2 coating is biocompatible and a good candidate material used as implants.展开更多
Immediate-early genes(IEGs) have long been used to visualize neural activations induced by sensory and behavioral stimuli. Recent advances in imaging techniques have made it possible to use endogenous IEG signals to v...Immediate-early genes(IEGs) have long been used to visualize neural activations induced by sensory and behavioral stimuli. Recent advances in imaging techniques have made it possible to use endogenous IEG signals to visualize and discriminate neural ensembles activated by multiple stimuli, and to map whole-brain-scale neural activation at single-neuron resolution. In addition, a collection of IEG-dependent molecular tools has been developed that can be used to complement the labeling of endogenous IEG genes and, especially, to manipulate activated neural ensembles in order to reveal the circuits and mechanisms underlying different behaviors. Here, we review these techniques and tools in terms of their utility in studying functional neural circuits. In addition, we provide an experimental strategy to measure the signal-to-noise ratio of IEG-dependent molecular tools, for evaluating their suitability for investigating relevant circuits and behaviors.展开更多
文摘目的探讨Arc基因表达及甲基化与幼龄鼠和成年鼠空间记忆形成的相关性。方法应用Morris水迷宫对2月龄幼龄鼠和6月龄成年鼠进行定位航行训练,以判断其空间记忆形成情况。采用RT-PCR检测定位航行训练后大鼠海马组织Arc基因mRNA的转录水平,并对海马基因组DNA进行亚硫酸盐处理,采用甲基化特异性聚合酶链反应(Methylation specific polymerase chain reaction,MS-PCR)检测Arc基因的甲基化情况,并进行DNA甲基化序列分析。结果随着定位航行训练入水次序及训练天数的增加,幼龄鼠逃避潜伏期缩短(P<0.05);而成年鼠在训练当天随着入水次序的增加,逃避潜伏期缩短,但第2天逃避潜伏期不仅与入水次序有关,也随着放入点距离的增大,逃避潜伏期相对延长(P<0.05),至第3天逃避潜伏期与入水次序及距离均无关,形成稳定的记忆能力。成年鼠定位航行训练组海马组织Arc基因mRNA的转录水平显著高于正常对照组(P<0.01),且高于幼龄鼠(P<0.05)。与幼龄鼠正常对照组相比,幼龄鼠和成年鼠定位航行训练组第8和24位点均无甲基化,成年鼠定位航行训练组与正常对照组相比,甲基化位点无变化。结论幼龄鼠瞬时记忆能力接近成年鼠,但形成稳定记忆的能力弱于成年鼠。Arc启动子甲基化影响其mRNA的表达,而Arc基因mRNA的转录水平与大鼠空间记忆能力及年龄相关,进一步证实了甲基化参与调解学习和记忆。
基金supported by Shanghai Science and Technology R&D Fund(Grant No.0952nm04400)the National Natural Science Foundation of China(GrantNos.51071168and30973041)the Innovation Fund of SIC,CAS(Grant No.Y06ZC3130G)
文摘Silicon （Si） incorporated porous TiO2 coating （Si-TiO2） prepared on titanium （Ti） by micro-arc oxidation （MAO） technique was demonstrated to be cytocompatible in previous studies. In view of the potential clinical applications, a detailed in vitro study of the biological activity of Si-Ti02 coating, in terms of osteoblast （MC3T3-EI cells） morphology, proliferation, differentiation and mineralization was performed. Immunofluo- rescent staining indicated that cells seeded on the Si-TiO2 coating showed improved adhesion with developing mature cytoskeletons, which contained numerous distinct and well-defined actin stress fibers in the cell mem- branes compared with those on the Ti02 coating and Ti plate. Results from proliferation assay showed that the proliferation rate of cells seeded on the Si-TiO2coating was significantly faster than that on the TiO2 coating and Ti plate. Furthermore, the analysis of osteogenic gene expression demonstrated that the Si-Ti02 coating stimulated the expression of osteoblast-related genes and promoted differentiation and mineralization of MC3T3-EI cells. In addition, the Si-TiO2 coating differentially regulated Wnt signaling pathway by up-regulating the expression of low-density lipoprotein （LDL） receptor-related protein 5 （LrpS）, and downregulating the expression of Dickkopf-1 （Dkkl）. All together, these results indicate that the investigated titanium with Si-TiO2 coating is biocompatible and a good candidate material used as implants.
基金supported by the National Natural Science Foundation of China(81571335,91432108 and81527901)grants from the Ministry of Science and Technology of China(2016YFA0501000)
文摘Immediate-early genes(IEGs) have long been used to visualize neural activations induced by sensory and behavioral stimuli. Recent advances in imaging techniques have made it possible to use endogenous IEG signals to visualize and discriminate neural ensembles activated by multiple stimuli, and to map whole-brain-scale neural activation at single-neuron resolution. In addition, a collection of IEG-dependent molecular tools has been developed that can be used to complement the labeling of endogenous IEG genes and, especially, to manipulate activated neural ensembles in order to reveal the circuits and mechanisms underlying different behaviors. Here, we review these techniques and tools in terms of their utility in studying functional neural circuits. In addition, we provide an experimental strategy to measure the signal-to-noise ratio of IEG-dependent molecular tools, for evaluating their suitability for investigating relevant circuits and behaviors.
