Effects of desferoxamine-induced hypoxia on neuronal human mu-opioid receptor gene expression.
Effects of desferoxamine-induced hypoxia on neuronal human mu-opioid receptor gene expression.
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DOI：
10.1016/j.bbrc.2010.06.032
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发表时间：
2010-07-16
影响因子：
3.1
通讯作者：
Ko, Jane L.
中科院分区：
文献类型：
--
作者：
Cook, Ryan J.;Karch, Christopher;Nahar, Pranjal;Rivera, Aida;Ko, Jane L.
用去铁胺（DFO）诱导的NMB细胞缺氧对神经元人μ阿片受体（hMOR）基因表达的影响。DFO以剂量和时间依赖性方式降低细胞活力并增加细胞谷胱甘肽水平。使用膜联蛋白-V-荧光素和碘化丙啶染色的共聚焦分析显示，在DFO攻击下存活/附着的细胞在形态上类似于对照（媒介物处理的）细胞。RT-PCR结果显示，缺氧诱导因子-1 α（HIF-1α）mRNA在存活神经元中表达增强。DFO处理8小时或更长时间下调hMOR信息，但不下调δ-阿片受体的信息。荧光素酶报告基因功能分析表明，hMOR 5′调控区（-357bp至翻译起始位点+1）含有活性启动子和抑制区（-422至-357bp）。与对照相比，DFO降低hMOR启动子活性。突变分析表明存在dsDNA和ssDNA元件，位于hMOR的富含CT的区域，介导DFO反应。RT-PCR进一步显示DFO对hMOR mRNA的稳定性没有影响。总之，DFO诱导的缺氧特异性地影响神经元hMOR基因在转录水平的表达，而不是转录后水平。
The effect of desferoxamine (DFO)-induced hypoxia on neuronal human mu-opioid receptor (hMOR) gene expression was investigated using NMB cells. DFO decreased cell viability and increased cellular glutathione levels in a dose- and time-dependent manner. Confocal analysis using annexin-V-fluorescein and propidium iodide staining revealed that surviving/attached cells under DFO challenge were morphologically similar to control (vehicle-treated) cells. RT-PCR analysis demonstrated that the hypoxia inducible factor-1α (HIF-1α) mRNA level was augmented in these surviving neurons. DFO treatment for 8hrs or longer down-regulated the hMOR message, but not that of the delta-opioid receptor. Functional analysis using luciferase reporter assay showed that the hMOR 5′-regulatory region, from −357bp to translational initiation site (+1), contains the active promoter with an inhibitory region located in the −422 to −357bp region. DFO decreased hMOR promoter activity as compared to control. Mutation analysis suggested the existence of both dsDNA and ssDNA elements, located in a CT-rich region of hMOR, mediating the DFO-response. RT-PCR further revealed that DFO exhibited no effect on hMOR mRNA stability. In conclusion, DFO-induced hypoxia specifically affects neuronal hMOR gene expression at the transcriptional, not post-transcriptional, level.
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DOI：
10.1016/s0169-328x(02)00357-1
发表时间：
2002-08-15
期刊：
MOLECULAR BRAIN RESEARCH
影响因子：
--
作者：
Ko, JL;Chen, HC;Loh, HH
通讯作者：
Loh, HH
影响因子：
4.7
作者：
Guo S;Miyake M;Liu KJ;Shi H
通讯作者：
Shi H
影响因子：
2.9
作者：
BAUMHAKER, Y;BENDOR, T;SARNE, Y
通讯作者：
SARNE, Y
影响因子：
4.7
作者：
Wendel, B;Hoehe, MR
通讯作者：
Hoehe, MR
DOI：
10.1073/pnas.0611704104
发表时间：
2007-03-06
影响因子：
11.1
作者：
Formisano, Luigi;Noh, Kyung-Min;Zukin, R. Suzanne
通讯作者：
Zukin, R. Suzanne