New therapy for cerebral infarction targeting of the control of mutant ubiquitin 1 (UBB+1) producted by RNA editing

以RNA编辑产生的突变泛素1（UBB 1）为靶点的脑梗塞新疗法

基本信息

批准号：
17590897
负责人：
URABE Takao
金额：
$ 2.24万
依托单位：
Juntendo University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2005
资助国家：
日本
起止时间：
2005 至 2006
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-17590897/
关键词：
RNA editing Cerebral infarction Mutant ubiquitin Proteasome Neuronal death Neuroprotection

项目摘要

Background and Purpose of this studyAccumulation of mutant ubiquitin-B (UBB^<+1>) produced by RNA editing in neurons is considered the hallmark of proteasomal dysfunction in neurodegenerative disorders, however no such evidence in ischemic brain has been reported. We investigated the contribution of UBB^<+1> in delayed neuronal death after transient global ischemia.Serial changes in UBB^<+1> protein and transcripts in gerbil brainWe evaluated the expression of UBB^<+1> transcripts and protein in gerbil brain after transient global ischemia. To investigate the serial change and cellular localization of UBB^<+1> protein, we performed immunohistochemical study in a gerbil brain after transient global ischemia. In the CA1 region, UBB^<+1> immunoreactivity appeared in the cytoplasm of pyramidal cells at 30 min post-ischemia, and the density of these neurons increased at day 2 and further increased at day 4 post-ischemia. In contrast, UBB^<+1> immunoreactivity was only transiently detected f … More rom 30 min to 1 day post-ischemia in CA3, dentate gyrus, and frontal cortex, but disappeared at day 2 post-ischemia.To investigate the serial change and cellular localization of UBB^<+1> mRNA, we performed RT-PCR study in a same model. UBB^<+1> mRNA was detected by RT-PCR in every region of the hippocampus and frontal cortex of ischemic gerbils and even in the non-ischemic control animals, and its expression level was independent of brain region and time after ischemia.Our results indicate induction and selective accumulation of UBB^<+1> protein in dying neurons of the CA1 region and suggest that UBB^<+1> is a potential mediator of ubiquitin proteasomal dysfunction and may have an important role in modifying delayed neuronal death.Evaluation of proteasomal function in vitro hypoxia modelTo assess the relationship of proteasomal function for ischemic neuronal death, we performed immunochemical study for hypoxia-inducible factor-1 using hypoxic culture system of neurons or /and astrocytes. Less

本研究的背景和目的神经元中RNA编辑产生的突变体泛素-B（UBB^+1）的积累被认为是神经退行性疾病中蛋白酶体功能障碍的标志，但在缺血性脑中尚未有此类证据的报道。 UBB^<+1> 在短暂性全局缺血后迟发性神经元死亡中的贡献。沙鼠脑中 UBB^<+1> 蛋白和转录物的系列变化我们评估了短暂性整体缺血后沙鼠脑中的UBB^+1 转录物和蛋白质为了研究UBB^+1 蛋白质的连续变化和细胞定位，我们在短暂性整体缺血后沙鼠脑中进行了免疫组织化学研究。缺血后30分钟，UBB^<+1>免疫反应性出现在锥体细胞的细胞质中，这些神经元的密度在第2天增加，并在第4天进一步增加相反，在缺血后 30 分钟至 1 天，CA3、齿状回和额叶皮质中仅短暂检测到 UBB^<+1> 免疫反应性，但在缺血后第 2 天消失。为了研究UBB^<+1> mRNA的序列变化和细胞定位，我们在同一模型中进行了RT-PCR研究，通过RT-PCR在每个区域检测到UBB^<+1> mRNA。缺血沙鼠甚至非缺血对照动物的海马和额叶皮层中都有表达，并且其表达水平与缺血后的脑区域和时间无关。我们的结果表明UBB^<+1>蛋白在垂死神经元中诱导和选择性积累的CA1区，表明UBB^+1是泛素蛋白酶体功能障碍的潜在介质，并且可能在改变迟发性神经元死亡中具有重要作用。蛋白酶体的评估体外缺氧模型的功能为了评估蛋白酶体功能与缺血性神经元死亡的关系，我们使用神经元或/和星形胶质细胞的缺氧培养系统进行了缺氧诱导因子-1的免疫化学研究。