[Molecular dissection of TDP-43 in ALS and FTLD].

Proteomic and immunochemical analyses have shown that hyperphosphorylated TDP-43 is a major component of ubiquitin-positive inclusions from brain of frontotemporal lobar degeneration (FTLD) patients. In 2008, TDP-43 gene mutations were discovered in familial and sporadic amyotrophic lateral sclerosis (ALS), indicating that TDP-43 protein abnormality is associated with neurodegeneration. We raised antibodies against 36 synthetic phosphopeptides and demonstrated that abnormal phosphorylation takes place in the C-terminal region of TDP-43. One antibody, pS409/410, stained the inclusions in both FTLD and ALS brains, with no nuclear staining. Immunoblotting revealed the presence of hyperphosphorylated 45 kDa band, smearing substances and 18-26 kDa fragments in deposits, and the band patterns were different between FTLD and ALS. Overexpression of TDP-43 C-terminal fragments as GFP-fusions resulted in formation of inclusions positive for antibodies to phosphorylated TDP-43 and ubiquitin. We further investigated the protease-resistant TDP-43 and found that it is also different between ALS and FTLD, supporting the idea that the different band patterns reflect different conformations of abnormal TDP-43. Interestingly, the C-terminal band pattern is indistinguishable among brain regions and spinal cord in each individual patient. These results suggest that abnormal TDP-43 produced in a cell may be transferred to different regions and propagated during disease progression.