Phosphorylation of S409/410 of TDP-43 is a consistent feature in all sporadic and familial forms of TDP-43 proteinopathies.
Phosphorylation of S409/410 of TDP-43 is a consistent feature in all sporadic and familial forms of TDP-43 proteinopathies.
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DOI：
10.1007/s00401-008-0477-9
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发表时间：
2009-02
影响因子：
12.7
通讯作者：
Lee VM
中科院分区：
文献类型：
--
作者：
Neumann M;Kwong LK;Lee EB;Kremmer E;Flatley A;Xu Y;Forman MS;Troost D;Kretzschmar HA;Trojanowski JQ;Lee VM
过度磷酸化、泛素化和N端截短的TAR DNA结合蛋白（TDP-43）的积累是大多数家族性和散发性FTLD-U和ALS形式的病理标志性病变，可归入TDP-43蛋白病。为了更深入地了解异常磷酸化在疾病过程中的作用，必须鉴定特异性磷酸化位点并产生磷酸化特异性抗体。在这里，我们开发和表征了新的大鼠单克隆抗体（1D 3和7A 9）提出的磷酸化的TDP-43的S409/410。这些抗体被用于通过免疫组织化学和生化分析研究64例伴有或不伴有运动神经元疾病的FTLD-U病例的大系列中S409/410磷酸化的存在，所述FTLD-U病例包括具有颗粒蛋白前体突变（n=5）、含缬沙汀蛋白（n=4）和与染色体9 p连锁（n=4）的家族性病例，18例ALS病例以及伴随TDP-43病理的其他神经退行性疾病（n=5）。我们的数据表明，TDP-43的S409/410的磷酸化是TDP-43蛋白病的散发性和家族性形式的整个谱中的病理性包涵体中高度一致的特征。通过免疫组织化学和免疫印迹分析，用这些mAb检测不到生理核TDP-43。虽然在FTLD-U和ALS的皮质脑区域中磷酸化C-末端片段的积累是一个强有力的发现，通常比磷酸化全长TDP-43条带丰富得多，但脊髓样品显示全长TDP-43在C-末端片段上占优势。这证明了皮质细胞与脊髓细胞内含物中不同的TDP-43物质组成。总体而言，这些mAb是高度特异性检测疾病相关异常TDP-43种类的有力工具，并且对于TDP-43蛋白病的神经病理学常规诊断以及TDP-43蛋白病的新兴细胞和动物模型的研究将是非常有用的。
Accumulation of hyperphosphorylated, ubiquitinated and N-terminally truncated TAR DNA-binding protein (TDP-43) is the pathological hallmark lesion in most familial and sporadic forms of FTLD-U and ALS, which can be subsumed as TDP-43 proteinopathies. In order to get more insight into the role of abnormal phosphorylation in the disease process, the identification of specific phosphorylation sites and the generation of phosphorylation-specific antibodies are mandatory. Here, we developed and characterized novel rat monoclonal antibodies (1D3 and 7A9) raised against phosphorylated S409/410 of TDP-43. These antibodies were used to study the presence of S409/410 phosphorylation by immunohistochemistry and biochemical analysis in a large series of 64 FTLD-U cases with or without motor neuron disease including familial cases with mutations in progranulin (n=5), valosin-containing protein (n=4) and linkage to chromosome 9p (n=4), 18 ALS cases as well as other neurodegenerative diseases with concomitant TDP-43 pathology (n=5). Our data demonstrate, that phosphorylation of S409/410 of TDP-43 is a highly consistent feature in pathologic inclusions in the whole spectrum of sporadic and familial forms of TDP-43 proteinopathies. Physiological nuclear TDP-43 was not detectable with these mAbs by immunohistochemistry and by immunoblot analyses. While the accumulation of phosphorylated C-terminal fragments was a robust finding in the cortical brain regions of FTLD-U and ALS, usually being much more abundant than the phsphorylated full-length TDP-43 band, spinal cord samples revealed a predominance of full-length TDP-43 over C-terminal fragments. This argues for a distinct TDP-43 species composition in inclusions in cortical versus spinal cord cells. Overall, these mAbs are powerful tools for the highly specific detection of disease-associated abnormal TDP-43 species and will be extremely useful for the neuropathological routine diagnostics of TDP-43 proteinopathies and for the investigation of emerging cellular and animal models for TDP-43 proteinopathies.
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