Molecular Dissection of TDP-43 Proteinopathies
Molecular Dissection of TDP-43 Proteinopathies
复制标题
DOI：
10.1007/s12031-011-9571-x
复制
发表时间：
2011-11-01
影响因子：
3.1
通讯作者：
Akiyama, Haruhiko
中科院分区：
文献类型：
--
作者：
Hasegawa, Masato;Nonaka, Takashi;Akiyama, Haruhiko
TDP-43已被鉴定为具有泛素阳性包涵体的额颞叶变性（FTLD-U）和肌萎缩侧索硬化（ALS）中的泛素阳性tau阴性胞质包涵体的主要组分。我们提出了代表人TDP-43的64个候选磷酸化位点中的36个的磷酸肽的抗体，并表明针对pS379、pS403/404、pS409、pS410和pS409/410的抗体标记了包涵体，但不标记核。免疫印迹分析表明，抗体识别TDP-43在类似的45 kDa，涂抹物质和18-26 kDa的C-末端片段。此外，C-末端片段的带型在神经病理亚型之间不同，但在每个个体患者的脑区域和脊髓之间无法区分。肌氨酸不溶性TDP-43的蛋白酶处理表明，C-末端片段的不同带型反映了疾病之间异常TDP-43分子的不同构象。这些结果表明，异常TDP-43的分子种类在疾病之间是不同的，并且它们在疾病进展期间从受影响的细胞传播到其他细胞，并决定疾病的临床病理表型。
TDP-43 has been identified as a major component of ubiquitin-positive tau-negative cytoplasmic inclusions in frontotemporal lobar degeneration with ubiquitin-positive inclusions (FTLD-U) and in amyotrophic lateral sclerosis (ALS). We raised antibodies to phosphopeptides representing 36 out of 64 candidate phosphorylation sites of human TDP-43 and showed that the antibodies to pS379, pS403/404, pS409, pS410 and pS409/410 labeled the inclusions, but not the nuclei. Immunoblot analyses demonstrated that the antibodies recognized TDP-43 at similar to 45 kDa, smearing substances and 18-26 kDa C-terminal fragments. Furthermore, the band patterns of the C-terminal fragments differed between neuropathological subtypes, but were indistinguishable between brain regions and spinal cord in each individual patient. Protease treatment of Sarkosyl-insoluble TDP-43 suggests that the different band patterns of the C-terminal fragments reflect different conformations of abnormal TDP-43 molecules between the diseases. These results suggest that molecular species of abnormal TDP-43 are different between the diseases and that they propagate from affected cells to other cells during disease progression and determine the clinicopathological phenotypes of the diseases.