Phosphorylated TDP-43 in frontotemporal lobar degeneration and amyotrophic lateral sclerosis

被引:589
作者
Hasegawa, Masato [1 ]
Ara, Tetsuaki [2 ]
Nonaka, Takashi [1 ]
Kametani, Fuyuki [1 ]
Yoshida, Mari [3 ]
Hashizume, Yoshio [3 ]
Beach, Thomas G. [4 ]
Buratti, Emanuele [5 ]
Baralle, Francisco [5 ]
Morita, Mitsuya [6 ]
Nakano, Imaharu [6 ]
Oda, Tatsuro [7 ]
Tsuchiya, Kuniaki [8 ]
Akiyama, Haruhiko [2 ]
机构
[1] Tokyo Metropolitan Org Med Res, Tokyo Inst Psychiat, Dept Mol Neurobiol, Setagaya Ku, Tokyo 1568585, Japan
[2] Tokyo Metropolitan Org Med Res, Tokyo Inst Psychiat, Dept Psychogeriatr, Tokyo 1568585, Japan
[3] Aichi Med Univ, Inst Med Sci Aging, Dept Neuropathol, Nagakute, Aichi 48011, Japan
[4] Sun Hlth Res Inst, Sun City, AZ USA
[5] Int Ctr Genet Engn & Biotechnol, I-34012 Trieste, Italy
[6] Jichi Med Univ, Dept Neurol, Shimotsuke Shi, Tochigi, Japan
[7] Natl Shimofusa Mental Hosp, Dept Neuropsychiat, Chiba, Japan
[8] Tokyo Metropolitan Matsuzawa Hosp, Dept Lab Med & Pathol, Setagaya Ku, Tokyo, Japan
关键词
D O I
10.1002/ana.21425
中图分类号
R74 [神经病学与精神病学];
学科分类号
摘要
Objective: TAR DNA-binding protein of 43kDa (TDP-43) is deposited as cytoplasmic and intranuclear inclusions in brains of patients with frontotemporal lobar degeneration with ubiquitinated inclusions (FTLD-U) and amyotrophic lateral sclerosis (ALS). Previous studies reported that abnormal phosphorylation takes place in deposited TDP-43. The aim of this study was to identify the phosphorylation sites and responsible kinases, and to clarify the pathological significance of phosphorylation of TDP-43. Methods: We generated multiple antibodies specific to phosphorylated TDP-43 by immunizing phosphopeptides of TDP-43, and analyzed FTLD-U and ALS brains by immunohistochemistry, immunoelectron microscopy, and immunoblots. In addition, we performed investigations aimed at identifying the responsible kinases, and we assessed the effects of phosphorylation on TDP-43 oligomerization and fibrillization. Results: We identified multiple phosphorylation sites in carboxyl-terminal regions of deposited TDP-43. Phosphorylation-specific antibodies stained more inclusions than antibodies to ubiquitin and, unlike existing commercially available anti-TDP-43 antibodies, did not stain normal nuclei. Ultrastructurally, these antibodies labeled abnormal fibers of 15nm diameter and on immunoblots recognized hyperphosphorylated TDP-43 at 45kDa, with additional 18 to 26kDa fragments in sarkosyl-insoluble fractions from FTLD-U and ALS brains. The phosphorylated epitopes were generated by casein kinase-1 and -2, and phosphorylation led to increased oligomerization and fibrillization of TDP-43. Interpretation: These results suggest that phosphorylated TDP-43 is a major component of the inclusions, and that abnormal phosphorylation of TDP-43 is a critical step in the pathogenesis of FTLD-U and ALS. Phosphorylation-specific antibodies will be powerful tools for the investigation of these disorders.
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页码:60 / 70
页数:11
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