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转录组相关研究(一)

RNA专题

英文题目Cognitive rigidity and BDNF-mediated frontostriatal glutamate neuroadaptations during spontaneous nicotine withdrawal.

中文题目:认知强直与脑源性神经营养因子介导的额侧谷氨酸神经适应。

作者Cole RD

作者单位:Department of Psychology and Neuroscience Program, Temple University,

Philadelphia, PA, 19122, USA国宾夕法尼亚州费城坦普尔大学心理学和神经科学系

期刊Neuropsychopharmacology.   发表时间:2019-11     IF6.22

DOI: 10.1038/s41386-019-0574-6

文章类型:研究型

英文摘要:

Cognitive flexibility is the ability to switch strategic responses adaptively in  changing environments. Cognitive rigidity imposed by neural circuit adaptations during nicotine abstinence may foster maladaptive nicotine taking in addicts. We  systematically examined the effects of spontaneous withdrawal in mice exposed to  either nicotine (6.3 or 18 mg/kg/day) or saline for 14 days on cognitive flexibility using an operant strategy set-shifting task. Because frontostriatal circuits are critical for cognitive flexibility and brain-derived neurotrophic factor (BDNF) modulates glutamate plasticity in these circuits, we also explored  the effects of nicotine withdrawal on these neurochemical substrates. Mice undergoing nicotine withdrawal required more trials to attain strategy-switching  criterion. Error analysis show that animals withdrawn from both nicotine doses committed higher perseverative errors, which correlated with measures of anxiety.

However, animals treated with the higher nicotine dose also displayed more strategy maintenance errors that remained independent of negative affect. BDNF mRNA expression increased in the medial prefrontal cortex (mPFC)  followingnicotine withdrawal. Surprisingly, BDNF protein declined in mPFC but was elevated in dorsal striatum (DS). DS BDNF protein positively correlated with perseverative and maintenance errors, suggesting mPFC-DS overflow of BDNF during withdrawal. BDNF-evoked glutamate release and synapsin phosphorylation was attenuated within  DS synapses, but enhanced in the nucleus accumbens, suggesting a dichotomous role

of BDNF signaling in striatal regions. Taken together, these data suggest that spontaneous nicotine withdrawal impairs distinct components of cognitive set-shifting and these deficits may be linked to BDNF-mediated alterations in glutamate signaling dynamics in discrete frontostriatal circuits.

中文摘要:

认知灵活性是在变化的环境中自适应地转换策略反应的能力。在尼古丁戒断期间,神经回路的适应性所造成的认知僵化可能会导致瘾君子对尼古丁的不适应。我们采用操作性策略集转移任务,系统地研究了尼古丁(6.318毫克/千克/日)或生理盐水对14天的小鼠自发戒断的影响。因为额叶纹状体电路对于认知灵活性和脑源性神经营养因子是至关重要的。因子(BDNF)调节谷氨酸在这些电路中的可塑性,我们还探讨了尼古丁撤回对这些神经化学底物的影响。接受尼古丁戒断的老鼠需要更多的试验来实现策略转换。误差分析表明,两种尼古丁剂量的动物都有较高的持续性错误,这与焦虑的测量结果相关。然而,用较高尼古丁剂量治疗的动物也显示出了与维持消极影响无关的策略维持错误。尼古丁戒断后,内侧前额叶皮质BDNF.mRNA表达增加。令人惊讶的是,在mPFCBDNF蛋白下降,但在背纹状体(DS)中升高。DS-BDNF蛋白与持续性错误和维持性错误呈正相关,提示BDNF在停药过程中的mPFC-DS溢出,BDNF诱发的谷氨酸释放和突触蛋白磷酸化在DS突触内减弱,但在伏隔核增强,提示BDNF信号在纹状体区域的双重作用。总之,这些数据表明,自发的尼古丁戒断损害了认知的不同组成部分,这些缺陷可能与BDNF介导的离散额-体回路谷氨酸信号动力学的改变有关。

 

 

英文题目:N-WASP knockdown upregulates inflammatory cytokines expression in human gingival  fibroblasts.

中文题目:N-WASP基因敲除上调人牙龈成纤维细胞炎性细胞因子的表达。

作者:Wang Y

单位:Department of Periodontology, School and Hospital of Stomatology, Shandong

University 山东省口腔医学院牙周病学系

期刊:ARCHIVES OF ORAL BIOLOGY  发表时间:2019-11  IF1.88

DOI: 10.1016/j.archoralbio.2019.104605

文章类型:研究型

英文摘要:

OBJECTIVE: The neuronal wiskott-aldrich syndrome protein (N-WASP) is a member of  the wiskott-aldrich syndrome protein (WASP) family. N-WASP plays a vital role in  promoting cell migration, receptor signaling and immune inflammatory responses. This study aimed to observe the changes in the expression of inflammatory factors and involving pathways after N-WASP knockdown in human gingival fibroblasts (HGFs).

DESIGN: Gingival inflammatory condition of N-WASP knockout mice was evaluated by  H&E staining. N-WASP in HGFs was knockdown by siRNA and the best knockdown efficiency was determined by qRT-PCR and immunofluorescence. The mRNA levels of interleukin (IL)-6, IL-8, C-C motif ligand 2 (CCL2), superoxide dismutase 2 (SOD2) and prostaglandin endoperoxide synthase 2 (PTGS2) were evaluated by qRT-PCR after N-WASP knockdown with or without mitogen-activated protein kinase (MAPK) and nuclear factor-κB (NF-κB) inhibitors. The protein levels of IL-6, IL-8 and CCL2 were assessed by ELISA. Western blotting was used to detect the activation of NF-κB and MAPK signaling pathways.

RESULTS: Gingival tissue from N-WASP knockout mice exhibited an inflammatory reaction. The expression of IL-6, IL-8, CCL2, SOD2 and PTGS2 was significantly upregulated after N-WASP knockdown in HGFs for 6, 24 and 48 h, except for the SOD2 at 6 h. N-WASP knockdown significantly activated the signaling pathways of NF-κB and MAPK. The inhibitors of p65, p38, ERK and JNK clearly decreased IL-6, IL-8, CCL2, SOD2 and PTGS2 expression after N-WASP knockdown. CONCLUSION: These data indicated that N-WASP deficiency in HGFs increases the production of inflammatory cytokine and is regulated via NF-κB and MAPK  signaling pathways.

中文摘要:

目的:神经元wiskott-aldrich综合征蛋白(N-WASP)是wiskott-aldrich综合征蛋白(WASP)家族的一员。黄蜂在促进细胞迁移,受体信号与免疫炎症反应本研究旨在观察N-WASP基因敲除人牙龈成纤维细胞(HGFs)后炎症因子表达及相关通路的变化。设计:采用H&E染色法评价N-WASP基因敲除小鼠牙龈炎症状况。用siRNA基因敲除HGFs中的N-黄蜂,并用qRT-PCR和免疫荧光检测其效率。白细胞介素(IL- 6IL-8C-C基序配体2CL2)、超氧化物歧化酶2SOD2)和前列腺素内过氧化物合酶2PGGS2)的mRNA水平在Q-RT-PCR检测N-WASP敲除或不含丝裂原活化蛋白激酶(MAPK)和核因子-κBNF-κB)抑制剂后的表达水平。ELISA法检测IL-6IL-8CCL2蛋白水平。结果:N-WASP基因敲除小鼠牙龈组织呈炎症反应。HGFsN-WASP基因敲除62448小时后,IL-6IL-8CCL2SOD2PTGS2的表达显著上调,但6小时的SOD2基因敲除明显激活了NF-κBMAPK的信号通路。p65p38ERKJNK抑制剂明显降低N-WASP基因敲除后IL-6IL-8CCL2SOD2PTGS2的表达。结论:HGFsN-WASP的缺失增加了炎性细胞因子的产生,并通过NF-κBMAPK信号途径进行调控。

 

 

英文题目:Novel BDNF-regulatory microRNAs in neurodegenerative disorders pathogenesis: An in silico study.

中文题目:神经退行性疾病发病机制中新的BDNF调节microRNAs:一项矽肺研究。

作者:Khani-Habibabadi F

单位:Department of Genetics, Faculty of Biological Sciences, Tarbiat Modares

University, Tehran, Iran.伊朗德黑兰大学塔比亚特莫代雷斯生物科学学院遗传学系

期刊:COMPUTATIONAL BIOLOGY AND CHEMISTRY  发表时间:2019-11  IF: 1.74

DOI: 10.1016/j.compbiolchem.2019.107153

文章类型:研究型

英文摘要:

Brain-derived neurotrophic factor (BDNF) is a neurotrophic factor with various roles in the central nervous system neurogenesis, neuroprotection, and axonal guide. By attaching to Tropomyosin receptor kinase B (TrkB) receptor, this protein triggers downstream signaling pathways which lead to cellular growth, proliferation, survival, and neuroplasticity. Deregulation at mRNA level is involved in various central nervous system disorders including, Huntington, Alzheimer\'s, Multiple Sclerosis, and Amyotrophic Lateral Sclerosis diseases. Considering the importance of BDNF functions, deciphering the regulatory mechanisms controlling BDNF expression level could pave the way to develop more accurate and efficient treatments for neurological diseases. Among different regulatory systems, microRNAs (miRNAs) play prominent roles by targeting genes 3\' untranslated regions. In this study, 127 validated and bioinformatic-predicted miRNAs with potentially regulatory roles in BDNF expression were analyzed.  Various aspects of miRNAsö possible functions were assessed by bioinformatic online tools to find their potential regulatory functions in signaling pathways,  neurological disorders, expression of transcription factors and miRNAs sponge. Analyzed data led to introduce 5 newly reported miRNAs that could regulate BDNF  expression level. Finally, high throughput sequencing data from different brain regions and neurological disorders were analyzed to measure correlation of candidate miRNAs with BDNF level in experimental studies. In this study, a list of novel miRNAs with possible regulatory roles in BDNF expression level involving in different neurological disorders was introduced.

中文摘要:

脑源性神经营养因子(BDNF)是一种神经营养因子,在中枢神经系统的神经发生、神经保护和轴突形成中起着多种作用。通过连接原肌球蛋白受体激酶BTrkB)受体,该蛋白触发下游信号通路,从而导致细胞生长、增殖、存活和神经可塑性。在mRNA水平上的放松调控涉及多种中枢神经系统疾病,包括亨廷顿、阿尔茨海默氏症、多发性硬化症和肌萎缩性侧索硬化症。考虑到BDNF功能的重要性,破译调控机制。控制BDNF表达水平的机制可以为开发更准确和更有效的BDNF奠定基础神经系统疾病的有效治疗。在不同的调控系统中,microRNAsmiRNAs)通过靶向基因3\'非翻译区发挥着重要作用。在这项研究中,我们分析了127个经验证和生物信息学预测的miRNAs,它们在BDNF表达中具有潜在的调节作用。利用生物信息学在线工具评估miRNAs的各个方面的可能功能,以发现它们在信号传导途径、神经系统疾病中的潜在调节功能,转录因子和miRNAs海绵的表达。分析数据,引入5个新报道的miRNAs,它们可以调节BDNF的表达水平。最后,对来自不同脑区和神经系统疾病的高通量测序数据进行分析,以测量实验研究中候选miRNAsBDNF水平的相关性。在这项研究中,我们介绍了一系列新的miRNAs,这些miRNAs可能在不同神经系统疾病中参与BDNF表达水平的调节。

 

 

英文题目:Evaluation of a co-extraction kit for mRNA, miRNA and DNA methylation-based body  fluid identification.

中文题目:一种基于mRNAmiRNADNA甲基化的联合提取试剂盒的评价。

作者:Watanabe K

单位:National Research Institute of Police Science, Chiba 277-0882, Japan. Electronic address: k-watanabe@nrips.go.jp.(国立警察科学研究所)

期刊:Leg Med      发表时间:2019-11       IF0.152

DOI: 10.1016/j.legalmed.2019.101630

文章类型:研究型

英文摘要:

Recently, messenger RNA (mRNA), micro RNA (miRNA), and DNA methylation (DNAm) have been reported as novel markers for body fluid identification (BFID). Comprehensive analysis of these markers should be a flexible and reliable BFID method for various types of forensic samples. However, independent extraction of  all targets can be difficult depending on the usable amounts of samples. In this  study, the applicability of a co-extraction kit for these molecules, the AllPrep DNA/RNA/miRNA Universal Kit (APU), was evaluated by comparing RNA and DNA extracted from blood and saliva stains by the APU with those extracted by standard kits for each molecule and by previously reported methods for mRNA/DNA or miRNA/DNA co-extraction. Electrophoresis using the Bioanalyzer platform and real-time PCR analysis revealed that the APU performed almost equivalently to each standard kit in the quality of RNA or DNA extracted and extraction efficiency of mRNAs, miRNAs, and DNA. Moreover, the APU outperformed the co-extraction methods, especially in RNA integrity and miRNA extraction efficiency. In addition, pyrosequencing revealed that the methylation ratios of DNA extracted by the APU were not different from those extracted by standard DNA  extraction kits. Overall, the APU is applicable to comprehensive analysis of mRNA/miRNA/DNAm markers for BFID analysis. Because the DNA eluate can also be used for DNA typing, the APU may be among the best choices for forensic examination of body fluid samples in terms of its flexibility and reliability in  BFID and efficiency in sample consumption.

中文摘要:

近来,信使RNAmRNA)、微RNAmiRNA)、DNA甲基化(DNAM)等已被报道为体液鉴定的新标志物,对这些标志物的综合分析应该是一种灵活、可靠的BFID方法。但是,根据可用的样本量,独立提取所有目标可能很困难。在这项研究中,通过比较从血液和唾液中提取的RNADNA,以及从每个分子的标准试剂盒和以前报道的mRNA/DNAmiRNA/DNA共提取的方法,评估了AllPrep.DNA/RNA/miRNA通用试剂盒(APU)对这些分子的适用性。利用生物分析平台进行电泳和实时PCR分析,结果表明APURNADNA的提取质量、mRNAsmiRNAsDNA的提取效率等方面几乎与标准试剂盒相当。此外,APURNA完整性和miRNA提取效率方面均优于联合提取法。此外,焦磷酸测序显示,用APU提取的DNA甲基化率与用标准DNA提取试剂盒提取的DNA甲基化率没有差异。总之,APU适用于BFID分析中mRNA/miRNA/DNAm标记的综合分析。由于DNA洗脱液也可用于DNA分型,APU可能是法医学的最佳选择。体液样品在其BFID和样品消耗效率方面的灵活性和可靠性方面的检查。

 

 

英文题目:Label-Free proteomic analysis reveals the differentiation between unfertilized and fertilized Beijing-You chicken eggs.

中文题目:无标记蛋白质组学分析揭示了未受精和受精的京优鸡蛋之间的区别。

作者:Zhang L

单位:Institute of Food Science and Technology, Chinese Academy of Agricultural

Sciences, Beijing 100193, PR China.中国农业科学院食品科学技术研究所

期刊:Int J Biol Macromol. 发表时间:2019-11    IF 2.858

DOI: 10.1016/j.ijbiomac.2019.10.189

文章类型:研究型

英文摘要:

Egg fertilization is a dynamic process, including varieties of biochemical changes. To better understand the molecular mechanisms during the egg embryo development, the objective of this study was to quantify protein expression changes between fertilized and unfertilized Beijing-You chicken eggs using label-free liquid chromatography-tandem mass spectrometry method. The results showed that a total of 1241 proteins were identified from fertilized and unfertilized eggs, 229 proteins were observed difference in fertilized eggs (p<0.05) compared with that in unfertilized eggs. The expressions of 86 proteins  were up-regulated and 48 proteins were down-regulated in fertilized eggs. STRING database analysis and Gene Ontology analysis results showed that these differentially expressed proteins significantly interacted and were involved in lipid transport and inflammatory response biological processes. The mRNA and protein expression levels of most differentially expressed proteins Apolipoprotein B, Fibrinogen alpha chain, Transferrin receptor protein 1, Phospholipid transfer protein and Vimentin were validated by RT-PCR and western blot. These results could provide possible novel insights for the molecular mechanism of egg fertilization.

中文摘要:

卵子受精是一个动态过程,包括各种生化变化。为了更好地了解鸡蛋胚胎发育过程中的分子机制,本研究采用无标记液相色谱-串联质谱法对北京油鸡受精卵和未受精卵的蛋白质表达进行了定量研究。结果表明,受精卵和未受精卵共鉴定出1241种蛋白,其中229种蛋白与未受精卵比较差异显著(p<0.05)。受精卵中86种蛋白表达上调,48种蛋白表达下调。数据库分析和基因本体分析结果表明,这些差异表达的蛋白质显著地相互作用,并参与脂质转运和炎症反应的生物学过程。用RT-PCRwestern.blot方法检测各组差异表达蛋白、载脂蛋白B、纤维蛋白原α链、转铁蛋白受体蛋白1、磷脂转移蛋白和波形蛋白的mRNA和蛋白表达水平。这些结果为卵子受精的分子机制提供了可能的新见解。

 

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