title: Molecular dissection of TDP-43 proteinopathies.
abstract: 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 ~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.
authors.affiliation: Department of Neuropathology and Cell Biology, Tokyo Metropolitan Institute of Medical Science, 2-1-6 Kamikitazawa, Setagaya-ku, Tokyo, 156-8506, Japan. hasegawa-ms@igakuken.or.jp
authors.forename: Masato
authors.initials: M
authors.lastname: Hasegawa
authors.forename: Takashi
authors.initials: T
authors.lastname: Nonaka
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Tsuji
authors.forename: Akira
authors.initials: A
authors.lastname: Tamaoka
authors.forename: Makiko
authors.initials: M
authors.lastname: Yamashita
authors.forename: Fuyuki
authors.initials: F
authors.lastname: Kametani
authors.forename: Mari
authors.initials: M
authors.lastname: Yoshida
authors.forename: Tetsuaki
authors.initials: T
authors.lastname: Arai
authors.forename: Haruhiko
authors.initials: H
authors.lastname: Akiyama
mesh_terms: D000690:Amyotrophic Lateral Sclerosis / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D000906:Antibodies / Q000378:metabolism; D001921:Brain / Q000378:metabolism / Q000473:pathology; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D057174:Frontotemporal Lobar Degeneration / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D006801:Humans; D015151:Immunoblotting; D045586:Intranuclear Inclusion Bodies / Q000378:metabolism / Q000473:pathology; D010446:Peptide Fragments / Q000235:genetics / Q000378:metabolism; D010766:Phosphorylation; D011487:Protein Conformation; D057177:TDP-43 Proteinopathies / Q000235:genetics / Q000378:metabolism* / Q000473:pathology*
chemical_list: D000906:Antibodies; D004268:DNA-Binding Proteins; D010446:Peptide Fragments