Progranulin deficiency leads to enhanced cell vulnerability and TDP-43 translocation in primary neuronal cultures
Progranulin deficiency leads to enhanced cell vulnerability and TDP-43 translocation in primary neuronal cultures
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DOI：
10.1016/j.brainres.2010.09.099
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发表时间：
2010-12-17
期刊：
影响因子：
2.9
通讯作者：
Jia, William
中科院分区：
文献类型：
--
作者：
Guo, Aobo;Tapia, Lucia;Jia, William
颗粒体蛋白前体基因 (PGRN) 的无效突变已被确定为具有泛素化包涵体的额颞叶痴呆的主要原因。在这种疾病中，一种通常位于核的 RNA 加工蛋白（称为 TAR DNA 结合蛋白 (TDP-43)）的泛素化聚集蛋白内含物在神经元细胞质 (FTLD-TDP) 中积累。为了确定这种临床病理学的各个方面是否可以在小鼠皮质神经元的原代培养物中建立，使用慢病毒载体引入小鼠PGRN-siRNA构建体来降低神经元培养物中的PGRN水平，并随后通过使用表达PGRN的慢病毒载体过度表达PGRN来挽救。 PGRN 的消耗增强了 caspase-3 的激活，并且 PGRN 缺陷的神经元表现出对正常亚致死剂量的 N-甲基-D-天冬氨酸 (NMDA) 和过氧化氢 (H2O2) 的脆弱性增强。 PGRN 缺陷神经元细胞质中的 TDP-43 蛋白水平相对于细胞核水平显着增加，这与 FTLD-TDP 患者大脑中的观察结果相似。我们的结果建立了 PGRN 缺陷的神经元培养模型，该模型显示了该疾病早期阶段的一些重要表型特征。研究结果进一步表明，这种额颞叶痴呆的种子可能在生命早期就已播下。 (C) 2010 年由 Elsevier B.V. 出版
Null mutations in the progranulin gene (PGRN) have been identified as a major cause of frontotemporal dementia with ubiquitinated inclusions. In this disorder, ubiquitinated, aggregated protein inclusions of a normally nuclear-located RNA processing protein called TAR DNA binding protein (TDP-43) accumulate in the neuronal cytoplasm (FTLD-TDP). To determine whether aspects of this clinical pathology can be established in primary cultures of mouse cortical neurons, PGRN levels were knocked down in neuronal cultures using lentiviral vectors to introduce mouse PGRN-siRNA constructs and subsequently rescued by overexpressing PGRN using a human PGRN-expressing lentiviral vector. The depletion of PGRN enhanced caspase-3 activation, and the PGRN-deficient neurons demonstrated enhanced vulnerability to normally sublethal doses of N-methyl-D-aspartic acid (NMDA) and hydrogen peroxide (H2O2). TDP-43 protein levels were markedly increased in the cytoplasm of PGRN-deficient neurons relative to nuclear levels, which is similar to observations in the brains of FTLD-TDP patients. Our results establish a neuronal culture model of the PGRN deficiency, which displays some of the important phenotypic characteristics of the early stages of the disease. The results further suggest that the seeds of this form of frontotemporal dementia may be sown early in life. (C) 2010 Published by Elsevier B.V.