Subcellular distribution of N-methyl-D-aspartic acid receptor subunit 1 in neural stem cells within subventricular zone of adult rats

The subcellular localization of N-methyI-D-aspartic acid receptor subunit 1 in neural stem cells of the subventricular zone of adult rats was detected using electron microscopy, following immunohistochemistry and immunogold-silver double staining. Results confirmed the presence of neural stem cells in the subventricular zone, which is a key neurogenic region in the central nervous system of adult mammals. The expression of N-methyI-D-aspartic acid receptor subunit 1 was higher than that of nestin and mainly distributed in the cell membrane, cytoplasm, rough endoplasmic reticulum and Golgi complex of neural stem cells.

泛癌分析揭示SREK1在低级别胶质瘤中促进CD274表达

目的剪接调节谷氨酸和富赖氨酸的蛋白质1(SREK1)在多种肿瘤中的泛癌分析,揭示SREK1在泛癌中的作用。方法利用在线数据库GEPIA 2、TIMER 2.0、TISIDB和cBioPortal分析SREK1表达对肿瘤患者预后的影响、在低级别胶质瘤(LGG)肿瘤组织中的表达、遗传变异的特征及其表达对肿瘤组织中免疫细胞的浸润和免疫—肿瘤靶基因的相关性分析。结果LGG肿瘤组织中,SREK1表达与记忆B细胞、活化的CD4+T细胞、Th2细胞、中性粒细胞、NKT细胞以及单核细胞和CD56dimNK细胞的浸润存在相关性(P均<0.05)。SREK1与免疫—肿瘤靶基因如信号传导及转录激活蛋白3(STAT3)、Ⅰ型干扰素受体1(IFNAR1)、核受体亚家族3C组成员1(NR3C1)和表皮生长因子受体(EGFR)、表面抗原分化簇274(CD274)等表达在LGG中均呈正相关(P均<0.05)。结论SREK1是LGG患者的危险因子之一,可能通过促进CD274的表达来加剧LGG的进展。

脂多糖对大鼠下丘脑室旁核GABA_BR1阳性神经元的激活作用

目的:探讨大鼠下丘脑室旁核(PVN)-氨基丁酸B受体亚单位1(GABABR1)阳性神经元对脂多糖(LPS)刺激的反应.方法:将大鼠腹腔注射LPS建立免疫应激模型,对照组注射等量的生理盐水,采用免疫荧光双标记与激光共聚焦显微镜技术,观察大鼠下丘脑室旁核内神经元Fos和GABABR1的标记情况以及它们在同一神经元内是否有共标记.结果:腹腔注射LPS可使大鼠PVN内表达Fos神经元数量显著增高,且PVN内部分神经元可同时被Fos和GABABR1双重标记,双重标记的神经元大约占Fos神经元的30%,占GAB-ABR1的24%.结论:下丘脑室旁核部分GABABR1阳性神经元参与了LPS诱导的免疫应激反应,它们可能在下丘脑-垂体-肾上腺轴的调节方面起重要作用.

孕期边缘型维生素A缺乏对新生鼠RARα、Src和NMDAR1表达的影响

目的探讨孕期边缘型维生素A缺乏(MVAD)对新生鼠海马中视黄酸核受体α(RARα)、肉瘤基因Src和N-甲基-D-天门冬氨酸受体1(NMDAR1)表达的影响。方法构建孕期维生素A正常(VAN)和MVAD大鼠模型(VAN组和MVAD组);分离培养新生鼠原代海马神经元,免疫荧光染色鉴定;采用real-time PCR和Western blotting检测新生鼠海马组织及体外培养的原代海马神经元中RARα、Src和NMDAR1的mRNA和蛋白表达;利用钙影像仪检测原代海马神经元钙兴奋性。结果免疫荧光染色显示,分离培养的新生鼠原代海马神经元95%表达神经元标志物神经元特异性烯醇化酶(NSE);MVAD组新生鼠海马组织和原代海马神经元中RARα、Src、NMDAR1的mRNA和蛋白表达水平明显低于VAN组(P<0.05);MVAD组新生鼠海马神经元的钙兴奋性明显低于VAN组(P<0.05)。结论孕期MVAD会降低海马RARα、Src和NMDAR1的表达,可能与孕期MVAD影响幼鼠的学习记忆功能有关。

囊泡膜谷氨酸转运体与ASIC3或TRPV1在大鼠结状神经节内的共存研究(英文)

为了探讨囊泡膜谷氨酸转运体(VGluTS)与酸敏感离子通道亚基3(ASIC3)或瞬时感受器电位香草酸亚型1(TRPV1)样阳性产物在大鼠迷走神经结状神经节(nodoseganglia,NG)内的分布和共存,本研究采用免疫荧光组织化学双重标记技术,在激光共聚焦显微镜下观察了大鼠NG内VGluT1或VGluT2分别与ASIC3或TRPV1的共存情况。结果显示:(1)在NG内,VGluT2、ASIC3和TRPV1主要见于中、小型神经元的胞浆内,大型神经元少见,而VGluT1阳性神经元数量较少,主要见于大型或中型神经元;(2)部分VGluT2阳性神经元同时显示ASIC3或TRPVl免疫活性,其中VGluT2/ASIC3双标神经元分别占VGluT2阳性神经元的43.06%,占ASIC3阳性神经元的58.74%;而VGluT2/TRPVl双标神经元分别占VGluT2或TRPVl阳性神经元的5617%和51.12%;(3)VGluTI与ASIC3或TRPV1双标神经元的数量很少,不到1%。以上结果提示,VGluT2主要存在于NG内中、小型神经元,这些神经元通常被认为是内脏伤害性信息的初级感受神经元,因而VGluT2分别与ASIC3或TRPVl的共存表明它们可能与内脏伤害性信息的产生和传递密切相关。

Monocyte chemotactic protein-inducing protein 1 negatively regulating asthmatic airway inflammation and mucus hypersecretion involvingγ-aminobutyric acid type A receptor signaling pathway in vivo and in vitro

Background:Mounting evidence,consistent with our previous study,showed thatγ-aminobutyric acid type A receptor(GABAAR)played an indispensable role in airway inflammation and mucus hypersecretion in asthma.Monocyte chemotactic protein-inducing protein 1(MCPIP1)was a key negative regulator of inflammation.Recent studies showed that inflammation was largely suppressed by enhanced MCPIP1 expression in many inflammatory diseases.However,the role and potential mechanism of MCPIP1 in airway inflammation and mucus hypersecretion in asthma were still not well studied.This study was to explore the role of MCPIP1 in asthmatic airway inflammation and mucus hypersecretion in both mice and BEAS-2B cells,and its potential mechanism.Methods:In vivo,mice were sensitized and challenged by ovalbumin(OVA)to induce asthma.Airway inflammation and mucus secretion were analyzed.In vitro,BEAS-2B cells were chosen.Interleukin(IL)-13 was used to stimulate inflammation and mucus hypersecretion in cells.MCPIP1 Lentiviral vector(LA-MCPIP1)and plasmid-MCPIP1 were used to up-regulate MCPIP1 in lung and cells,respectively.MCP-1,thymic stromal lymphopoietin(TSLP),mucin 5AC(MUC5AC),MCPIP1,and GABAARβ2 expressions were measured in both lung and BEAS-2B cells.Immunofluorescence staining was performed to observe the expression of GABAARβ2 in cells.Results:MCPIP1 was up-regulated by LA-MCPIP1(P<0.001)and plasmid-MCPIP1(P<0.001)in lung and cells,respectively.OVA-induced airway inflammation and mucus hypersecretion,OVA-enhanced MCP-1,TSLP,MUC5AC,and GABAARβ2 expressions,and OVA-reduced MCPIP1 were significantly blunted by LA-MCPIP1 in mice(all P<0.001).IL-13-enhanced MCP-1,TSLP,MUC5AC,and GABAARβ2 expressions,and IL-13-reduced MCPIP1 were markedly abrogated by plasmid-MCPIP1 in BEAS-2B cells(all P<0.001).Conclusion:The results of this study suggested that OVA and IL-13-induced airway inflammation and mucus hypersecretion were negatively regulated by MCPIP1 in both lung and BEAS-2B cells,involving GABAAR signaling pathway.

MicroRNA-502-3p regulates GABAergic synapse function in hippocampal neurons

Gamma-aminobutyric acid(GABA)ergic neurons,the most abundant inhibitory neurons in the human brain,have been found to be reduced in many neurological disorders,including Alzheimer's disease and Alzheimer's disease-related dementia.Our previous study identified the upregulation of microRNA-502-3p(miR-502-3p)and downregulation of GABA type A receptor subunitα-1 in Alzheimer's disease synapses.This study investigated a new molecular relationship between miR-502-3p and GABAergic synapse function.In vitro studies were perfo rmed using the mouse hippocampal neuronal cell line HT22 and miR-502-3p agomiRs and antagomiRs.In silico analysis identified multiple binding sites of miR-502-3p at GABA type A receptor subunitα-1 mRNA.Luciferase assay confirmed that miR-502-3p targets the GABA type A receptor subunitα-1 gene and suppresses the luciferase activity.Furthermore,quantitative reve rse transcription-polymerase chain reaction,miRNA in situ hybridization,immunoblotting,and immunostaining analysis confirmed that overexpression of miR-502-3p reduced the GABA type A receptor subunitα-1 level,while suppression of miR-502-3p increased the level of GABA type A receptor subunitα-1 protein.Notably,as a result of the overexpression of miR-502-3p,cell viability was found to be reduced,and the population of necrotic cells was found to be increased.The whole cell patch-clamp analysis of human-GABA receptor A-α1/β3/γ2L human embryonic kidney(HEK)recombinant cell line also showed that overexpression of miR-502-3p reduced the GABA current and overall GABA function,suggesting a negative correlation between miR-502-3p levels and GABAergic synapse function.Additionally,the levels of proteins associated with Alzheimer s disease were high with miR-502-3p overexpression and reduced with miR-502-3p suppression.The present study provides insight into the molecular mechanism of regulation of GABAergic synapses by miR-502-3p.We propose that micro-RNA,in particular miR-502-3p,could be a potential therapeutic to rget to modulate GABAergic synapse function in neurological disorders,including Alzheimer's disease and Alzheimer's diseaserelated dementia.

不同强度电针对PCPA失眠大鼠下丘脑γ-氨基丁酸及受体的影响

通过观察不同强度电针对睡眠节律紊乱大鼠下丘脑γ-氨基丁酸(GABA)及受体(GABRA1)的影响,初步探讨针灸治疗失眠的作用机制及不同强度电针的效应差异.用免疫组织化学技术观察失眠大鼠下丘脑GABA阳性细胞的表达情况,用逆转录-聚合酶链反应(RT-PCR)检测失眠大鼠下丘脑GABRA1 mRNA表达改变.研究发现,对氯苯丙氨酸(PCPA)化失眠大鼠下丘脑GABA阳性细胞染色较浅,且表达量减少,GABRA1mRNA表达明显降低(P<0.01);经电针治疗5 d后,失眠大鼠下丘脑GABA阳性神经元染色较深,表达量较多,GABRA1 mRNA表达显著升高(P<0.01),且2 V电针刺激作用比1 V电针刺激更为明显.结果表明电针可增加失眠大鼠下丘脑GABA及受体的表达,调节睡眠-觉醒周期,发挥镇静催眠作用,且2 V电针刺激效果优于1 V电针刺激.

电针对坐骨神经慢性缩窄损伤大鼠脊髓N-甲基-D-天冬氨酸受体表达的影响

目的：通过观察坐骨神经慢性缩窄损伤（CCI）大鼠脊髓相应节段N-甲基-D-天冬氨酸（NMDA）受体表达的变化和电针干预对其的影响，探讨电针干预神经病理性痛的脊髓机制。方法：清洁级雄性SD大鼠，随机分为3组，每组20只。假手术组仅分离坐骨神经，但不进行结扎；模型组采用CCI法制备神经病理性痛模型；电针组于CCI术后11～17d应用韩氏穴位神经刺激仪进行电针干预，针灸美容针刺入大鼠损伤侧“委中”穴与“环跳”穴，刺激时间a0rain，1次jd。免疫组化法测定大鼠脊髓NMDA受体2B亚基（NR2B）的表达；WesterIlblot法测定大鼠脊髓NMDA受体1亚基（NR1）和NR2B蛋白的表达；逆转录一聚合酶链反应法测定大鼠脊髓NR1mRNA和NR2B1TIRNA的表达。结果：与假手术组比较，模型组脊髓NR1蛋白及其mRNA表达量均升高（P〈0．01，P〈0．05）；与模型组比较，电针组脊髓NR1蛋白及其mRNA表达均被逆转（均P〈0．05）。各组脊髓NR2B蛋白及其mRNA的表达差异均无统计学意义（P〉0．05）。结论：电针减轻大鼠神经病理性痛的机制之一，可能与有效地下调脊髓NR1的功能有关。