Biochemical and histopathological alterations in TAR DNA-binding protein-43 after acute ischemic stroke in rats
Biochemical and histopathological alterations in TAR DNA-binding protein-43 after acute ischemic stroke in rats
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DOI：
10.1111/j.1471-4159.2010.06860.x
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发表时间：
2011-03-01
影响因子：
4.7
通讯作者：
Shimohata, Takayoshi
中科院分区：
文献类型：
--
作者：
Kanazawa, Masato;Kakita, Akiyoshi;Shimohata, Takayoshi
核因子TAR DNA结合蛋白-43（TDP-43）被认为通过其蛋白水解切割、异常磷酸化、亚细胞再分布和不溶解产生TDP-43阳性神经元胞内包涵体而在人类神经退行性疾病（所谓TDP-43蛋白病）的发病机制中起作用。本研究的目的是阐明TDP-43对急性缺血性卒中的特异性生化和组织病理学变化。成年雄性大鼠进行90分钟的大脑中动脉闭塞。我们研究了蛋白水解裂解，磷酸化，亚细胞定位和溶解度的TDP-43的免疫印迹和组织病理学检查使用缺血和假手术的皮质。急性缺血性卒中后，TDP-43全长（43 kDa）的水平下降，25 kDa的C-末端片段增加，这可以解释为TDP-43的蛋白水解裂解。急性缺血性卒中后观察到TDP-43的胞浆再分布和核分布改变，而未观察到TDP-43的异常磷酸化和不溶解以及细胞内包涵体的形成。TDP-43胞质再分布的缺血神经元表达泛素和激活的caspase 3，末端脱氧核苷酸转移酶介导的尿苷5 '-三磷酸-生物素缺口末端标记阳性。总之，TDP-43的生化和组织病理学改变在大鼠急性缺血性卒中后被确定，虽然有非常小的相似性TDP-43的变化在急性缺血性卒中和TDP-43蛋白质病变中观察到的。
P>Nuclear factor TAR DNA-binding protein-43 (TDP-43) is considered to play roles in pathogenesis of human neurodegenerative diseases, so-called TDP-43 proteinopathy, via its proteolytic cleavage, abnormal phosphorylation, subcellular redistribution, and insolubilization generating TDP-43-positive neuronal intracellular inclusions. The purpose of this study was to elucidate biochemical and histopathological alternations in TDP-43 specific to acute ischemic stroke. Adult male rats were subjected to a 90-min middle cerebral artery occlusion. We examined the proteolytic cleavage, phosphorylation, subcellular localization, and solubility of TDP-43 by immunoblottings and histopathological examinations using the ischemic and sham-operated cortex. The level of full-length TDP-43 (43 kDa) decreased and that of the 25-kDa C-terminal fragment increased after acute ischemic stroke, which can be explained by proteolytic cleavage of TDP-43. Cytoplasmic redistribution and altered nuclear distribution of TDP-43 was observed after acute ischemic stroke, whereas abnormal phosphorylation and insolubilization of TDP-43 as well as formation of intracellular inclusions were not observed. Ischemic neurons with the cytoplasmic redistribution of TDP-43 expressed ubiquitin and activated caspase 3 and were terminal deoxynucleotidyl transferase-mediated uridine 5'-triphosphate-biotin nick end labeling-positive. In conclusion, biochemical and histopathological alterations in TDP-43 were identified in rats after acute ischemic stroke, although there was very less similarity between TDP-43 alterations observed in acute ischemic stroke and those observed in TDP-43 proteinopathy.