Molecular Brain Research最新文献_第6页

Tetracycline-regulated gene expression in the NSC-34-tTA cell line for investigation of motor neuron diseases 四环素调控NSC-34-tTA细胞系基因表达在运动神经元疾病研究中的应用

Molecular Brain Research Pub Date : 2005-10-31 DOI: 10.1016/j.molbrainres.2005.07.010

Elisabetta Babetto , Alessandra Mangolini , Milena Rizzardini , Monica Lupi , Laura Conforti , Paola Rusmini , Angelo Poletti , Lavinia Cantoni

{"title":"Tetracycline-regulated gene expression in the NSC-34-tTA cell line for investigation of motor neuron diseases","authors":"Elisabetta Babetto , Alessandra Mangolini , Milena Rizzardini , Monica Lupi , Laura Conforti , Paola Rusmini , Angelo Poletti , Lavinia Cantoni","doi":"10.1016/j.molbrainres.2005.07.010","DOIUrl":"10.1016/j.molbrainres.2005.07.010","url":null,"abstract":"<div>The motor neuron-like cell line NSC-34 has become a widely used in vitro model for motor neuron biology and pathology. We established a tetracycline-regulated gene expression system in this cell line by stably transfecting pTet-Off, which codifies for the tetracycline transactivator, the regulatory protein tTA. The monoclonal cell lines (NSC-34-tTA) were evaluated for the presence of functional tTA after transient transfection with pBI-EGFP, analyzing the expression of the reporter gene enhanced green fluorescent protein. We evaluated the regulation of tTA function with doxycycline using fluorescence microscopy and quantitative cytofluorimetric analysis on viable transfected cells. The best-regulated cell line (NSC-34-tTA40) had a 66.4-fold induction for the reporter gene fluorescence in comparison to NSC-34.Alpha-tubulin, GAP-43 and phosphorylated medium and heavy neurofilaments, proteins of importance for the motor neuronal phenotype, were evident in NSC-34-tTA40 by Western blot and immunocytochemistry; they were expressed similarly in NSC-34-tTA40 and in NSC-34.The cDNA of human Cu/Zn superoxide dismutase, a gene of interest for amyotrophic lateral sclerosis, was cloned into pBI-EGFP, downstream of the tetracycline-responsive bidirectional promoter. This plasmid was transiently transfected into NSC-34-tTA40, and the functionality of bidirectional transcription was verified by determining the expression of enhanced green fluorescent protein and of human Cu/Zn superoxide dismutase. Both proteins were regulated by doxycycline.This novel cell line, NSC-34 tTA40, that permits tetracycline-regulated gene expression may prove useful to unravel the mechanisms of motor neuron degeneration.</div>","PeriodicalId":100932,"journal":{"name":"Molecular Brain Research","volume":"140 1","pages":"Pages 63-72"},"PeriodicalIF":0.0,"publicationDate":"2005-10-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.molbrainres.2005.07.010","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"25270509","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 15

LPS induces stefin A3 expression in mouse primary cultured glial cells LPS诱导小鼠原代培养的胶质细胞中stefin A3的表达

Molecular Brain Research Pub Date : 2005-10-31 DOI: 10.1016/j.molbrainres.2005.07.007

Toru Hosoi , Sawako Suzuki , Yasunobu Okuma , Akiko Kawakami , Naoki Ogawa , Koichiro Ozawa , Yasuyuki Nomura

引用次数: 4

Identification of novel differentially expressed genes in human astrocytomas by cDNA representational difference analysis 利用cDNA代表性差异分析鉴定人类星形细胞瘤中新的差异表达基因

Molecular Brain Research Pub Date : 2005-10-31 DOI: 10.1016/j.molbrainres.2005.06.015

Sueli M. Oba-Shinjo , Mario H. Bengtson , Sheila M.B. Winnischofer , Christian Colin , Cleber G. Vedoy , Zizi de Mendonça , Suely K.N. Marie , Mari C. Sogayar

{"title":"Identification of novel differentially expressed genes in human astrocytomas by cDNA representational difference analysis","authors":"Sueli M. Oba-Shinjo , Mario H. Bengtson , Sheila M.B. Winnischofer , Christian Colin , Cleber G. Vedoy , Zizi de Mendonça , Suely K.N. Marie , Mari C. Sogayar","doi":"10.1016/j.molbrainres.2005.06.015","DOIUrl":"10.1016/j.molbrainres.2005.06.015","url":null,"abstract":"<div>Diffuse infiltrating gliomas are the most common tumors of the central nervous system (CNS), naturally progressing from a lower-grade to a higher-grade malignancy. Several genetic alterations have been correlated with astrocytic tumors; however, a number of as yet unknown genes may also be involved. Therefore, we set out to search for genes that are differentially expressed in anaplastic astrocytoma and normal CNS tissue by applying a PCR-based subtractive hybridization approach, namely, representational difference analysis (RDA). The results of DNA sequencing of a sample (96 cDNA clones) from the subtracted library allowed the identification of 18 different genes, some of which were represented by several cDNA clones, coding for the Np95, LMO1, FCGBP, DSCAM, and taxilin proteins. Quantitative real-time PCR analysis for five of these genes was performed using samples of astrocytic tumors of different grades, confirming their higher expression when compared to non-tumoral CNS tissue. Identification of differentially expressed genes present in gliomas but not in normal CNS tissue is important not only to better understand the molecular basis of these cancers, but also to generate diagnostic DNA chips, which may be useful in future therapeutic intervention.</div>","PeriodicalId":100932,"journal":{"name":"Molecular Brain Research","volume":"140 1","pages":"Pages 25-33"},"PeriodicalIF":0.0,"publicationDate":"2005-10-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.molbrainres.2005.06.015","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"24943701","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 46

NAPOR-3 RNA binding protein is required for apoptosis in hippocampus 海马细胞凋亡需要NAPOR-3 RNA结合蛋白

Molecular Brain Research Pub Date : 2005-10-31 DOI: 10.1016/j.molbrainres.2005.07.006

Alessandra Pacini , Annarita Toscano , Valentina Cesati , Andrea Cozzi , Elena Meli , Lorenzo Di Cesare Mannelli , Ferdinando Paternostro , Paolo Pacini , Domenico E. Pellegrini-Giampietro

{"title":"NAPOR-3 RNA binding protein is required for apoptosis in hippocampus","authors":"Alessandra Pacini , Annarita Toscano , Valentina Cesati , Andrea Cozzi , Elena Meli , Lorenzo Di Cesare Mannelli , Ferdinando Paternostro , Paolo Pacini , Domenico E. Pellegrini-Giampietro","doi":"10.1016/j.molbrainres.2005.07.006","DOIUrl":"10.1016/j.molbrainres.2005.07.006","url":null,"abstract":"<div>NAPOR-3 is a central nervous system RNA binding protein that is associated with downstream mRNA targets and has been demonstrated to be selectively overexpressed during apoptotic cell death. In this study, we first examined the regional distribution of NAPOR-3 mRNA in the adult rat brain by in situ hybridization: the transcript was abundantly expressed in many brain regions, mostly in gray matter, including the CA1–CA4 regions and dentate gyrus of the hippocampus, the piriform cortex and the cerebellar granule cell layer. We then investigated the role of NAPOR-3 in neuronal cell death by monitoring its mRNA and protein expression levels using semiquantitative RT-PCR and Western blotting, respectively. NAPOR-3 was overexpressed in rat organotypic slices exposed to staurosporine and to oxygen–glucose deprivation (OGD), an in vitro model of apoptotic cerebral ischemia, but not when exposed to glutamate toxicity. Our results also demonstrate that NAPOR-3 gene overexpression is an early step in the chain of signaling events leading to apoptosis, taking place upstream of caspase-3 activation. Finally, antisense-mediated downregulation of NAPOR-3 gene expression protected hippocampal cultures against OGD-induced apoptosis and prevented caspase-3 activation. Our results demonstrate that NAPOR-3 gene overexpression is necessary for the execution of OGD-induced programmed cell death.</div>","PeriodicalId":100932,"journal":{"name":"Molecular Brain Research","volume":"140 1","pages":"Pages 34-44"},"PeriodicalIF":0.0,"publicationDate":"2005-10-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.molbrainres.2005.07.006","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"25243842","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 8

High intrinsic oxidative stress may underlie selective vulnerability of the hippocampal CA1 region 高内在氧化应激可能是海马CA1区选择性易感性的基础

Molecular Brain Research Pub Date : 2005-10-31 DOI: 10.1016/j.molbrainres.2005.07.018

Xinkun Wang , Ranu Pal , Xue-wen Chen , Nanteetip Limpeanchob , Keshava N. Kumar , Elias K. Michaelis

引用次数: 99

Regional expression of brain derived neurotrophic factor (BDNF) is correlated with dynamic patterns of promoter methylation in the developing mouse forebrain 脑源性神经营养因子(BDNF)的区域表达与发育中的小鼠前脑启动子甲基化的动态模式相关

Molecular Brain Research Pub Date : 2005-10-31 DOI: 10.1016/j.molbrainres.2005.06.014

Kathleen E. Dennis, Pat Levitt

{"title":"Regional expression of brain derived neurotrophic factor (BDNF) is correlated with dynamic patterns of promoter methylation in the developing mouse forebrain","authors":"Kathleen E. Dennis, Pat Levitt","doi":"10.1016/j.molbrainres.2005.06.014","DOIUrl":"10.1016/j.molbrainres.2005.06.014","url":null,"abstract":"<div>Brain Derived Neurotrophic Factor (BDNF) plays an important role in brain development and plasticity. BDNF gene expression is known to be dynamically regulated during development, but the regulatory controls of normal differential expression are not well understood. Methylation of CpG dinucleotides within gene promoters is emerging as an important epigenetic control mechanism of transcription, and the BDNF complex promoter contains several CpG dinucleotides. We determined BDNF expression in the developing mouse forebrain and examined whether there were correlated patterns of methylation at CpG dinucleotides within the BDNF promoter. The data show that BDNF is dynamically expressed in the mouse forebrain and that expression is correlated with differential methylation specifically at CpG dinucleotides in eIV of the mouse BDNF promoter. These studies demonstrate that DNA methylation of this regulatory region may be an important mechanism controlling differential expression of BDNF during forebrain development.</div>","PeriodicalId":100932,"journal":{"name":"Molecular Brain Research","volume":"140 1","pages":"Pages 1-9"},"PeriodicalIF":0.0,"publicationDate":"2005-10-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.molbrainres.2005.06.014","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"25221829","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 65

Stress induces changes of internal standard genes of amygdala 应激引起杏仁核内部标准基因的改变

Molecular Brain Research Pub Date : 2005-10-31 DOI: 10.1016/j.molbrainres.2005.07.003

Shigeru Maeda , Ichiro Nakatsuka , Takuya Miyawaki , Takuo Kuboki , Masahiko Shimada

引用次数: 2

Inhibitory effect of green tea extract on β-amyloid-induced PC12 cell death by inhibition of the activation of NF-κB and ERK/p38 MAP kinase pathway through antioxidant mechanisms 绿茶提取物通过抗氧化机制抑制NF-κB和ERK/p38 MAP激酶通路的激活，从而抑制β-淀粉样蛋白诱导的PC12细胞死亡

Molecular Brain Research Pub Date : 2005-10-31 DOI: 10.1016/j.molbrainres.2005.07.009

Sun Young Lee , Jae Woong Lee , Heesoon Lee , Han Soo Yoo , Yeo Pyo Yun , Ki Wan Oh , Tae Youl Ha , Jin Tae Hong

{"title":"Inhibitory effect of green tea extract on β-amyloid-induced PC12 cell death by inhibition of the activation of NF-κB and ERK/p38 MAP kinase pathway through antioxidant mechanisms","authors":"Sun Young Lee , Jae Woong Lee , Heesoon Lee , Han Soo Yoo , Yeo Pyo Yun , Ki Wan Oh , Tae Youl Ha , Jin Tae Hong","doi":"10.1016/j.molbrainres.2005.07.009","DOIUrl":"10.1016/j.molbrainres.2005.07.009","url":null,"abstract":"<div>Beta-amyloid peptide (Aβ) is considered responsible for the pathogenesis of Alzheimer's disease (AD). Several lines of evidence support that Aβ-induced cytotoxicity is mediated through the generation of reactive oxygen species (ROS). Thus, agents that scavenge ROS level may usefully impede the development or progress of AD. Green tea extract has been known to have such antioxidant properties. Our previous studies demonstrate that green tea extract protected ischemia/reperfusion-induced brain cell death by scavenging oxidative damages of macromolecules. In this study, we investigated the effects of green tea extract on Aβ-induced oxidative cell death in cultured rat pheochromocytoma (PC12) cells. PC12 cells treated with Aβ25–35 (10–50 μM) showed intracellular ROS elevation, the formation of 8-oxodG (an oxidized form of DNA), and underwent apoptotic cell death in a dose-dependent manner. Aβ25–35 treatment upregulated pro-apoptotic p53 at the gene level, and Bax and caspase-3 at the protein level, but downregulated anti-apoptotic Bcl-2 protein. Interestingly, co-treated green tea extract (10–50 μg/ml) dose-dependently attenuated Aβ25–35 (50 μM)-induced cell death, intracellular ROS levels, and 8-oxodG formation, in addition to p53, Bax, and caspase-3 expression, but upregulated Bcl-2. Furthermore, green tea extract prevented the Aβ25–35-induced activations of the NF-κB and ERK and p38 MAP kinase pathways. Our study suggests that green tea extract may usefully prevent or retard the development and progression of AD.</div>","PeriodicalId":100932,"journal":{"name":"Molecular Brain Research","volume":"140 1","pages":"Pages 45-54"},"PeriodicalIF":0.0,"publicationDate":"2005-10-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.molbrainres.2005.07.009","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"25002127","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 99

Naloxone's suppression of spontaneous and food-conditioned locomotor activity is diminished in mice lacking either the dopamine D2 receptor or enkephalin 在缺乏多巴胺D2受体或脑啡肽的小鼠中，纳洛酮对自发性和食物条件运动活动的抑制作用减弱

Molecular Brain Research Pub Date : 2005-10-31 DOI: 10.1016/j.molbrainres.2005.07.016

Michael D. Hayward , Malcolm J. Low

{"title":"Naloxone's suppression of spontaneous and food-conditioned locomotor activity is diminished in mice lacking either the dopamine D2 receptor or enkephalin","authors":"Michael D. Hayward , Malcolm J. Low","doi":"10.1016/j.molbrainres.2005.07.016","DOIUrl":"10.1016/j.molbrainres.2005.07.016","url":null,"abstract":"<div>Mice lacking the D2 dopamine receptor (D2−/−) and congenic to the C57BL/6J background were tested for opioid-mediated locomotor activity to examine the involvement of the D2 dopamine receptor in opioid pharmacology. Morphine-stimulated locomotor activity did not significantly differ between the two genotypes. The opioid antagonist naloxone dose-dependently decreased spontaneous motor activity in wild-type mice but was without significant effect in D2−/− mice. The magnitude of food-conditioned increases in locomotor activity in wild-type mice and D2−/− mice was similar but naloxone did not decrease conditioned motor activity in D2−/− mice. Spontaneous locomotor activity of mice lacking the endogenous opioids β-endorphin and/or enkephalin was also tested and we found that naloxone did not reduce activity in mice specifically lacking enkephalin. We suggest that the D2 dopamine receptor is necessary for modulation of spontaneous locomotor activity stimulated by the endogenous opioid enkephalin.</div>","PeriodicalId":100932,"journal":{"name":"Molecular Brain Research","volume":"140 1","pages":"Pages 91-98"},"PeriodicalIF":0.0,"publicationDate":"2005-10-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.molbrainres.2005.07.016","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"25271064","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 14

Free colour illustrations on the web 免费彩色插图在网络上

Molecular Brain Research Pub Date : 2005-10-31 DOI: 10.1016/S0169-328X(05)00383-9

引用次数: 0